Scholarly article on topic 'Pyridoacridine alkaloids from deep-water marine organisms: Structural elucidation'

Pyridoacridine alkaloids from deep-water marine organisms: Structural elucidation Academic research paper on "Chemical sciences"

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Abstract of research paper on Chemical sciences, author of scientific article — Sabrin R.M. Ibrahim, Gamal A. Mohamed

Abstract Pyridoacridine alkaloids are unique marine nitrogenous compounds that represent a large family of alkaloids. They have been reported from different marine organisms like sponges, ascidians, anemones, prosobranch mollusk, and tunicates. Attention to pyridoacridines has risen because of their significant biological activities. The present review emphasizes mainly on pyridoacridines isolated marine organisms over the last years. Thus, the synthetic ones were not discussed. Herein, 95 pyridoacridine alkaloids isolated from marine organisms have been retrieved, in addition to their classification, isolation, sources, structures, molecular weight, physical, and (UV, IR, 1H and 13C NMR) spectral data.

Academic research paper on topic "Pyridoacridine alkaloids from deep-water marine organisms: Structural elucidation"

Bulletin of Faculty of Pharmacy, Cairo University (2016) xxx, xxx-xxx

Cairo University

Faculty of Pharmacy

Cairo University Bulletin of Faculty of Pharmacy, Cairo University

www.elsevier.com/locate/bfopcu www.sciencedirect.com

BULLETIN OF FACULTY OF PHARMACY,

CAIRO UNIVERSITY

REVIEW PAPER

Pyridoacridine alkaloids from deep-water marine organisms: Structural elucidation

Sabrin R.M. Ibrahim a,b*, Gamal A. Mohamedc,d

a Department of Pharmacognosy and Pharmaceutical Chemistry, College of Pharmacy, Taibah University, Al Madinah Al Munawwarah 30078, Saudi Arabia

b Department of Pharmacognosy, Faculty of Pharmacy, Assiut University, Assiut 71526, Egypt

c Department of Natural Products and Alternative Medicine, Faculty of Pharmacy, King Abdulaziz University, Jeddah 21589, Saudi Arabia

d Department of Pharmacognosy, Faculty of Pharmacy, Al-Azhar University, Assiut Branch, Assiut 71524, Egypt

Received 22 June 2016; revised 24 August 2016; accepted 31 August 2016

KEYWORDS

Alkaloids; Pyridoacridine; Marine organisms; NMR spectral data

Abstract Pyridoacridine alkaloids are unique marine nitrogenous compounds that represent a large family of alkaloids. They have been reported from different marine organisms like sponges, ascidians, anemones, prosobranch mollusk, and tunicates. Attention to pyridoacridines has risen because of their significant biological activities. The present review emphasizes mainly on pyri-doacridines isolated marine organisms over the last years. Thus, the synthetic ones were not discussed. Herein, 95 pyridoacridine alkaloids isolated from marine organisms have been retrieved, in addition to their classification, isolation, sources, structures, molecular weight, physical, and (UV, IR, 1H and 13C NMR) spectral data.

© 2016 Published by Elsevier B.V. on behalf of Faculty of Pharmacy, Cairo University. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

Contents

1. Introduction........................................................................................................................................................00

2. Isolation and structural characterization of pyridoacridines......................................................................................00

2.1. Ultraviolet visible spectroscopy (UV)..............................................................................................................00

2.2. Infrared spectroscopy (IR)..............................................................................................................................00

2.3. NMR spectroscopy........................................................................................................................................00

2.4. Mass spectroscopy (MS)................................................................................................................................00

* Corresponding author at: Department of Pharmacognosy and Pharmaceutical Chemistry, College of Pharmacy, Taibah University, Al Madinah Al Munawwarah 30078, Saudi Arabia. Fax: +966 581183034.

E-mail address: sabrinshaur@gmail.com (S.R.M. Ibrahim). Peer review under responsibility of Faculty of Pharmacy, Cairo University.

http://dx.doi.org/10.1016/j.bfopcu.2016.08.003

1110-0931 © 2016 Published by Elsevier B.V. on behalf of Faculty of Pharmacy, Cairo University.

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

2.5. Stereochemistry determination............................................................ 00

Conflict of interest......................................................................... 00

References............................................................................... 00

1. Introduction

Secondary metabolites from natural sources still provide potential drug candidates with unique skeletons that are interesting for many synthetic approaches. Chemistry researches of marine natural products have yielded great numbers of impor-

Figure 1 Basic skeletons of pyridoacridine alkaloids.

tant metabolites with significant bioactivities. Marine organisms yield various toxic metabolites in order to prevent parasitism and predation as well as to mediate spatial compe-tition.1'2 Among these toxic metabolites are pyridoacridines. They are the largest group of alkaloids isolated from marine organisms. They have been reported from sponges, ascidians, anemones' tunicates' and prosobranch mollusk' which are decorated with bright colors.3'4 They have different colors: yellow, deep red, orange, blue, or purple. Their colors were attributed to the presence of pyridoacridines. Pyridoacridines colors change according to the pH. So, they may be used as an indicator. This property is due to the presence of basic nitrogen in the pyridine ring that associated with a chromophore. Generally, pyridoacridines are crystalline compounds with melting points >300 0C. They were isolated as salts of hydrochloric acid. The optical activity of some pyridoacridines is due to the additional asymmetric side chain. They are planar poly-cyclic heteroaromatic compounds, having 11H-pyrido[4,3,2, mn]acridine (1) or 8H-pyrido[4,3,2-nm]acridone (2) skeletons (Fig. 1), usually possessing different alkylamine side chains

Table 1 Tetracyclic pyridoacridine alkaloids.

Pantherinine (1)

H2N 9 lis 7a N

Purple powder; IR (KBr) vmax: 3810 (br), 3720 (br), 1665 (w), 1630 (s) cm"1; UV (MeOH) kmax (e): 254 (16,110), 316 (3950), 472 (br, 1620) nm; 1H NMR (DMSO-d6, 300 MHz): dH 7.87 (d, J = 8.7 Hz, H-1), 7.96 (dd, J = 8.8, 1.8 Hz, H-2), 8.99 (d, J = 1.8 Hz, H-4), 8.97 (d, J = 5.5 Hz, H-5), 9.18 (d, J = 5.5 Hz, H-6), 6.58 (s, H-10), 6.60, 8.46 (NH); 'H NMR (CDCl3/CD3OD, 300 MHz): dH 7.82 (d, J = 8.7 Hz, H-1), 7.89 (dd, J = 8.7, 2.1 Hz, H-2), 8.72 (d, J =2.1 Hz, H-4), 8.69 (d, J = 5.7 Hz, H-5), 9.10 (d, J = 5.7 Hz, H-6), 6.64 (s, H-10); South Australia ascidian Aplidium pantherinum34

Pantherinine acetate (2)

Cystodytin A (3)

111 N 1 I1al 2 ^ 3 4

18 16 N 12 10a[| OCb, Lib 'SS.

13 II I5

1 O 19 3 O 8 7a ^ 6 N 7

Red glass: UV (MeOH) kmax (e): 255 (5794), 293 (2811), 432 (1369) nm; IR (KBr) mmax: 3340, 1700 (w), 1655 (s), 1521 (s) cm"1; 'H NMR (CDCl3, 500 MHz): dH 8.12 (d, J = 8.5 Hz, H-1), 7.98 (dd, J = 8.5, 2.0 Hz, H-2), 8.66 (d, J = 2.0 Hz, H-4), 8.55 (d, J = 5.5 Hz, H-5), 9.29 (d, J = 5.5 Hz, H-6), 8.82 (s, H-10), 8.50 (NH), 2.38 (CH3); 13C NMR (CDCl3, 125 MHz): dC 132.21 (C-1), 135.17 (C-2),

122.31 (C-3), 125.91 (C-4), 123.37 (C-4a), 136.01 (C-4b), 120.75 (C-5), 150.03 (C-6), 144.79 (C-7a),

178.32 (C-8) 151.34 (C-9), 121.78 (C-10), 137.06 (C-10a), 116.33 (C-10b), 144.91 (C-10c), 170.38 (Ac); LRMS m/z (%): 369 (25), 367 (24), 327 (96), 325 (100), 300 (63), 298 (67); South Australia ascidian Aplidium pantherinum34

Yellow crystals; mp 181-183 0C; UV (MeOH) kmax (e): 225 (35,000), 272 (25,000), 380 (11,400) nm; IR (KBr) vmax: 3290, 2925, 2850, 1660, 1640, 1590, 1520, 1330, 1300, 1180, 860, 760 cm"1; 1H NMR (CDCl3/CD3OD (2:1), 400 MHz): dH 8.07 (dd, J = 8.2, 1.4 Hz, H-1), 7.76 (ddd, J = 8.2, 8.1, 1.3 Hz, H-2), 7.64 (ddd, J = 8.1, 8.1, 1.4 Hz, H-3), 8.30 (dd, J = 8.1, 1.3 Hz, H-4), 8.22 (d, J = 5.5 Hz, H-5), 8.81 (d, J = 5.5 Hz, H-6), 3.08 (t, J = 6.4 Hz, H-12), 3.59 (t, J = 6.4 Hz, H-13), 6.01 (brs, H-14), 5.50 (qq, J = 1.4, 1.3 Hz, H-16), 1.65 (d, J = 1.4 Hz, H-18), 1.93 (d, J = 1.3 Hz, H-19); 13C NMR (CDCl3/CD3OD (2:1), 100 MHz): dC 131.6 (C-1), 131.7 (C-2), 129.8 (C-3), 122.8 (C-4), 121.3 (C-4a), 136.9 (C-4b), 119.4 (C-5), 149.0 (c-6), 145.8 (C-7a), 183.2 (C-8), 132.0 (C-9), 152.4 (C-10), 149.8 (C-10a), 117.5 (c-10b), 145.0 (C-11a), 31.3 (C-12), 38.4 (C-13), 167.8 (C-15), 118.1 (C-16), 150.8 (C-17), 26.7 (C-18), 19.4 (C-19); EIMS m/z: 359 [M + 2] + , 357 [M] + , 328, 273, 260, 247; HRFABMS m/z: 360.1707 [M + H + 2H] + (calcd for C22H22O2N3 360.1712); Okinawan tunicate Cystodytes dellechiajei20

Table 1 (continued)

Cystodytin B (4)

Cystodytin C (5)

Cystodytin D (6)

Cystodytin E (7)

Yellow crystals; mp 181-183 °C; UV (MeOH) kmax (e): 225 (35,000), 272 (25,000), 380 (11,400) nm; IR (KBr) vmax: 3290, 2925, 2850, 1660, 1640, 1590, 1520, 1330, 1300, 1180, 860, 760 cm"1; 'H NMR (CDCl3/CD3OD (2:1), 400 MHz): dH 8.07 (dd, J = 8.2, 1.7 Hz, H-1'), 7.76 (ddd, J = 8.2, 8.1, 1.3 Hz, H-2'), 7.64 (ddd, J = 8.1, 8.1, 1.7 Hz, H-3'), 8.34 (dd, J = 8.1, 1.3 Hz, H-4'), 8.33 (d, J = 5.5 Hz, H-5'), 8.89 (d, J = 5.5 Hz, H-6'), 6.67 (s, H-9'), 3.11 (t, J = 6.4 Hz, H-12'), 3.60 (t, J = 6.4 Hz, H-13'), 6.19 (qq, J = 6.9, 1.5 Hz, H-17'), 1.60 (dq, J = 1.5, 1.2 Hz, H-18'), 1.51 (dq, J = 6.9, 1.2 Hz, H-19'); 13C NMR (CDCl3/CD3OD (2:1), 100 MHz): dC 131.3 (C-1'), 131.8 (C-2'), 129.8 (C-3'), 122.8 (C-4'), 121.4 (C-4'a), 137.0 (C-4'b), 119.5 (C-5'), 149.1 (C-6'), 183.2 (C-8'), 145.9 (C-7'a), 132.0 (C-9'), 152.5 (C-10'), 149.9 (C-10'a), 117.5 (C-10'b), 145.0 (C-11'a), 30.9 (C-12'), 39.3 (C-13'), 170.3 (C-15'), 130.3 (c-16'), 130.0 (c-17'), 11.9 (C-18'), 13.4 (C-19'); EIMS m/z: 359 [M + 2] + , 357 [M] + , 328, 273, 260, 247; HRFABMS m/z: 360.1707 [M + H + 2H] + (calcd for C22H22O2N3 360.1712); Okinawan tunicate Cystodytes dellechiajei20

(e): 228 (29,900), 272 (29,100), 380 (11,800) 1.

Light yellow crystals; mp 257-259°C; UV (MeOH) k nm; IR (KBr) vmax: 3400, 2930, 2850, 1660, 1640, 1580, 760 cm"1; *H NMR (CDCl3/CD3OD (2:1), 400 MHz): dH 8.25 (d, J = 8.2 Hz, H-1), 7.90 (dd, J = 8.2, 8.0 Hz, H-2), 7.81 (dd, J = 8.0, 8.0 Hz, H-3), 8.50 (d, J = 8.0 Hz, H-4), 8.44 (d, J = 5.5 Hz, H-5), 9.04 (d, J = 5.5 Hz, H-6), 6.86 (s, H-9), 3.25 (t, J = 6.4 Hz, H-12), 3.75 (t, J = 6.4 Hz, H-13), 2.29 (s, H-16), 1.19 (s, H-18, 19); 13CNMR(CDCl3/ CD3OD (2:1), 100 MHz): dC 131.7 (C-1, 2), 129.8 (C-3), 122.9 (C-4), 121.1 (C-4a), 137.2 (C-4b), 119.5 (C-5), 149.5 (C-6), 145.3 (C-7, 11a), 183.5 (C-8); 132.4 (C-9),153.2 (C-10), 149.9 (C-10a), 117.0 (C-10b), 31.4 (C-12), 38.6 (C-13), 172.7 (C-15), 47.6 (C-16), 65.0 (C-17), 28.9 (C-18), 28.9 (C-19); EIMS m/z: 377 [M + 2H] + , 375 [M] + , 373, 328, 315, 273, 259, 247; Okinawan tunicate Cystodytes

dellechiajei20

Brown-colored amorphous solids; [a]30D -160 (c 0.3, CHCl3); UV (MeOH) kmax (e): 214 (37,000), 274 (30,000), 384 (11,000) nm; IR (KBr) vmax: 3400, 2930, 2850, 1650 cm-1; *H NMR (DMSO-d6, 400 MHz): dH 8.24 (d, H-1), 7.94 (dd, H-2), 7.64 (dd, H-3), 8.94 (d, H-4), 8.39 (d, H-5), 9.10 (d, H-6), 7.20 (s, H-9), 5.56 (m, H-12), 4.16 (m, H-13A), 3.96 (m, H-13B), 6.07 (m, H-14), 5.62 (brs, H-16), 1.64 (brs, H-18), 2.16 (m, H-19); 13C NMR (DMSO-d6, 100 MHz): dC 131.7 (C-1), 131.9 (C-2), 129.9 (C-3), 123.0 (C-4), 121.8 (C-4a), 137.1 (C-4b), 119.2 (C-5), 150.1 (C-6), 144.9 (C-7a), 183.5 (c-8), 131.2 (C-9), 150.0 (C-10), 146.7 (C-10a), 117.4 (C-10b), 144.9 (C-11a), 71.2 (C-12), 41.6 (C-13), 169.3 (C-15), 117.4 (C-16), 153.4 (C-17), 27.3 (C-18), 20.0 (C-19); FABMS m/z: 376 [M + 2H] + , 358 [M-H2O + 2H] +; HRFABMS m/z: 376.1669 [M + 2H]+ (calcd for C22H22N3O3 376.1661); Okinawan marine tunicate Cystodytes dellecbiajei12

Brown-colored amorphous solids; [a]30D -160 (c 0.3, CHCl3); UV (MeOH) kmax (e): 214 (37,000), 274 (30,000), 384 (11,000) nm; IR (KBr) vmax: 3400, 2930, 2850, 1650 cm-1; *H NMR (DMSO-d6, 400 MHz): dH 6.36 (m, H-14), 6.48 (m, H-17), 1.82 (brs, H-18), 1.73 (dd, J = 6.9, 1.1 Hz, H-19); FABMS m/z: 376 [M + 2H] + , 358 [M-H2O + 2H] + ; HRFABMS m/z: 376.1669 [M + 2H]+ (calcd for C22H22N3O3 376.1661); Okinawan marine tunicate Cystodytes dellecbiajei12

Cystodytin F (8)

133 (c 0.3, CHCl3); UV (EtOH) km

(e): 225 (30,000), 272 11

Yellow-colored amorphous solids; [a]D0 (23,000), 380 (11,000) nm; IR (KBr) vmax: 3330, 2940, 2850, 1660, 1590, 1175, 1110 cm"1; *H NMR (cDCl3, 400 MHz): dH 7.92 (dt, J = 7.3, 1.0 Hz, H-2), 7.82 (dt, J = 8.3, 1.0 Hz, H-3), 8.59 (dd, J = 8.1, 1.8 Hz, H-4), 8.58 (d, J = 5.4 Hz, H-5), 9.25 (d, J = 5.4 Hz, H-6), 7.12 (s, H-9), 5.40 (t, J = 4.9 Hz, H-12), 3.99 (ddd, J = 14.0, 5.4, 3.9 Hz, H-13A), 3.82 (ddd, J = 14.0, 6.4, 5.4 Hz, H-13B), 5.86 (m, H-14), 5.57 (brs, H-16), 2.00 (d, J = l.0 Hz, H-18), 1.81 (d, J = 2.0 Hz, H-19), 3.50 (s, OCH3); 13C NMR (cDCl3, 100 MHz): dC 130.1 (C-1), 132.2 (C-2), 129.9 (C-3), 122.0 (C-4a), 137.4 (C-4b), 119.5 (C-5), 151.2 (C-6), 146.8 (C-7a), 183.8 (C-8), 131.7 (C-9), 150.1 (C-10), 149.7 (C-10a), 118.2 (C-10b), 145.4 (C-11a), 76.72 (C-12), 43.25 (C-13), 167.0 (C-15), 118.4 (C-16), 150.8 (C-17), 27.09 (c-18), 19.77 (C-19), 57.75 (OCH3); EIMS m/z: 389 [M + 2H] + , 357, 328, 302, 277, 262, 247; HREIMS m/z: 389.1749 [M + 2H]+ (calcd for C23H23O3N3 389.1740); Okinawan marine tunicate Cystodytes dellecbiajei12

(continued on next page)

Table 1 (continued)

Cystodytin G (9)

Yellow-colored amorphous solids; [a]Du -133 (c 0.3, CHCl3); UV (EtOH) kmax (e): 225 (30,000), 272 (23,000), 380 (11,000) nm; IR (KBr) vmax: 3330, 2940, 2850, 1660, 1590, 1175, 1110 cm-1; *H NMR (cDCl3, 400 MHz): dH 6.37 (dd, J = 6.8, 1.5 Hz, H-14), 6.24 (t, J = 4.9 Hz, H-17), 1.78 (t, J =1.2 Hz, H-18), 1.72 (dd, J = 6.8, 1.2 Hz, H-19), 3.52 (s, OCH3); 13C NMR (CDCl3, 100 MHz): dC 130.8 (C-17), 12.46 (C-18), 13.92 (C-19), 57.82 (OCH3); EIMS m/z: 389 [M + 2H] + , 357, 328, 302, 277, 262, 247; HREIMS m/z: 389.1749 [M + 2H]+ (calcd for C23H23O3N3 389.1740); Okinawan marine tunicate Cystodytes dellecbiajei12

Cystodytins H (10) and I (11)

Cystodytin J (12)

Cystodytin K (13)

H3CO >13 13 HN

Iminoquinone (14)

14 NH3 O2CCF3

Yellow-colored amorphous solids; [a]30D -29.1 (c 0.3, CHCl3); UV (EtOH) kmax (e): 225 (37,000), 273 (25,000), 382 (10,000) nm; IR (neat) vmax: 3320, 2910, 2840, 1730, 1650, 1590, 1540, 1250, 1190 cm-1; *H NMR (CDCl3, 400 MHz): dH 8.32 (brd, J = 8.3 Hz, H-l), 7.93 (dt, J = 7.8, l.5 Hz, H-2), 7.83 (brt, J = 7.3 Hz, H-3), 8.58 (dd, J = 8.1, 1.8 Hz, H-4), 8.57 (d, J = 5.4 Hz, H-5), 9.24 (d, J = 5.4 Hz, H-6), 6.99 (s, H-9), 6.85 (t, J = 3.4 Hz, H-12), 4.14 (m, H-13A), 3.94 (m, H-13B), 5.71 (m, H-14), 5.53 (brs, H-16), 5.35 (2H, m, -CH=CH-), 2.83 (2H, m, -CH=CH-CH2), 2.50 (2H, t, J = 6.3 Hz, H-2'), 2.04 (2H, m, -CH=CH-CH2), 1.79 (d, J = 1.5 Hz, H-18), 0.88 (t, J = 6.3 Hz, H-18'), 1.97 (d, J = 1.5 Hz, H-19), 1.70 (2H, m, CH2), 1.25 (22H, m, CH2); FABMS m/z: 640 [M + 2H] + , 374 [M + 2H-CigH33O2 + H] + , 358, 274, 263, 259; Okinawan marine tunicate Cystodytes dellecbiajei12

Yellow solid; 1H NMR (CDCl3, 500 MHz): dH 8.29 (d, J = 8.1 Hz, H-1), 7.94 (ddd, J = 8.1, 7.1, 1.4 Hz, H-2), 7.83 (ddd, J =8.1, 7.1, 1.4 Hz, H-3), 8.42 (d, J =8.1 Hz, H-4), 8.18 (d, J = 5.5 Hz, H-5), 8.94 (d, J = 5.5 Hz, H-6), 6.85 (s, H-9), 3.25 (t, J = 6.4 Hz, H-12), 3.79 (dt, J = 6.4, 5.9 Hz, H-13), 6.59 (s, H-14), 2.02 (s, H-16); 13C NMR (CDCl3, 125 MHz): dC 131.87 (C-1), 131.87 (C-2), 129.83 (C-3), 122.84 (C-4), 121.78 (C-4a), 136.92 (C-4b), 118.99 (C-5), 149.76 (C-6), 146.48 (C-7a), 183.31 (C-8), 132.82 (C-9), 152.17 (C-10), 150.33 (C-10a), 117.84 (C-10b), 145.32 (C-11a), 31.72 (C-12), 39.28 (C-13), 170.43 (C-15), 23.30 (C-16); FABMS (3-nitrobenzyl alcohol) m/z: 318 [M + H] + , 247; FABMS (glycerol) m/z: 320 [(M + 2) + H] + , 247; HRFABMS (glycerol) m/z: 320.1390 [(M + 2) + H] + (calcd for C19H18N3O2 320.1399); Fijian ascidian Cystodytes sp.35

Yellow solid; [a]D -292 (c 0.065, MeOH); UV-vis (MeOH/TFA) kmax (log e): 207 (4.57), 272 (4.17), 382 (3.67) nm; UV-Vis (MeOH/KOH) kmax (log e): 204 (4.46), 270 (4.17), 380 (3.71) nm; IR vmax: 3303, 2926, 1655, 1587, 1465, 1112, 1022 cm-1; 1H NMR (DMSO-d6, 300 MHz): dH 8.25 (d, J = 8.3 Hz, H-1), 8.03 (t, J = 8.3 Hz, H-2), 8.02 (t, obsc, H-3), 8.96 (d, J = 7.9 Hz, H-4), 9.06 (d, J = 5.4 Hz, H-5), 9.27 (d, J = 5.5 Hz, H-6), 6.84 (s, H-9), 5.28 (t, J = 4.3 Hz, H-12), 3.54 (m, H-13), 7.94 (obsc, H-14), 1.68 (s, H-2'), 3.42 (s, OCH3); 13C NMR (DMSO-d6, 75 MHz): dC 131.0 (C-1), 131.9 (C-2), 129.7 (C-3), 124.1 (C-4), 121.6 (C-4a), 137.3 (C-4b), 120.3 (C-5), 149.9 (C-6), 146.0 (C-7a), 132.0 (C-9), 150.5 (C-10), 146.2 (C-10a), 117.7 (C-10b), 144.5 (C-11a), 76.1 (C-12), 42.4 (C-13), 169.3 (C-1'), 22.3 (C-2'), 56.8 (OCH3); FABMS m/z: 350 [M + 2H + H] + ; HRFABMS m/z: 350.1507 [M + H]+ (calcd for C20H20N3O3 350.1505); Zealand ascidian Lissoclinum notti36

Brown solid; UV (MeOH-TFA) kmax (log e): 226 (3.97), 272 (3.95), 387 (3.49) nm; UV (MeOH-KOH) kmax (log e): 268 (3.97), 376 (3.49) nm; IR vmax (film): 3072, 1678, 1202, 1131 cm-1; 1H NMR (DMSO-d6, 500 MHz): dH 8.23 (d, J = 7.5 Hz, H-1), 8.03 (dd, J = 7.5, 7.5 Hz, H-2), 7.94 (m, H-3), 8.95 (d, J = 8.0 Hz, H-4), 9.05 (d, J = 5.0 Hz, H-5), 9.26 (d, J = 5.0 Hz, H-6), 7.11 (s, H-9), 5.67 (brm, H-12), 3.50 (m, H-13A), 3.18 (m, H-13B), 7.97 (m, H-14), 6.40 (brs, H-15); 13C NMR (DMSO-d6, 125 MHz): dC 130.7 (C-1), 132.0 (C-2), 130.0 (C-3), 124.1 (C-4), 121.7 (C-4a), 136.7 (C-4b), 120.4 (C-5), 150.0 (C-6), 146.3 (C-7a), 183.0 (C-8), 130.9 (C-9), 149.3 (C-10), 151.7 (C-10a), 117.8 (c-10b), 144.3 (C-11a), 64.2 (C-12), 44.3 (C-13); HRFABMS (glycerol) m/z: 294.1245 [M + 2H]+ (calcd for

C17H16N3O2 294.1243); FABMS (3-nitrobenzyl alcohol) m/z (rel. int.): 292 [M + H] + Indonesian ascidian Eusynstyela latericius37

(100);

Table 1 (continued)

Styelsamine A (15) cf3co2 h^ i /

cf3co2h

12 ho.

Styelsamine C (17) CF3C02H4*

W H O'^ H

Styelsamine D (18)

NH3O2CCF3

Diplamine (19)

15 NH3O2CCF3

Styelsamine B ( 16) 1 2

Purple solid; UV (MeOH-TFA) kmax (log e): 222 (sh) (3.86), 244 (3.71), 276 (3.93), 294 (3.96), 386 (3.17), 554 (3.02) nm; UV (MeOH-KOH) kmax (log e): 272 (3.85), 378 (3.39) nm; IR (KBr) vmax: 3102, 2942, 1678, 1584, 1443, 1202, 1143 cm-1; *H NMR (DMSO-d6, 500 MHz): dH 13.70 (brs, H-1), 8.33 (d, J = 6.0 Hz, H-2), 7.65 (d, J = 6.0 Hz, H-3), 8.24 (d, J = 8.5 Hz, H-4), 7.24 (dd, J = 8.0, 8.0 Hz, H-5), 7.69 (dd, J = 8.0, 8.0 Hz, H-6), 7.99 (d, J = 8.5 Hz, H-7), 11.13 (brs, H-8), 7.79 (s, H-10), 10.98 (brs, H-12), 5.62 (brd, J = 7.0 Hz, H-13), 3.07 (m, H-14A), 2.89 (m, H-14B), 8.18 (brs, H-15), 6.47 (brs, H-16); 13C NMR (DMSO-d6, 125 MHz): dC 143.6 (C-2), 105.5 (C-3), 149.1 (C-3a), 113.9 (C-3b), 125.4 (C-4), 122.6 (C-5), 134.8 (C-6), 117.9 (c-7), 140.9 (C-7a), 126.6 (C-8a), 118.3 (C-9), 117.4 (C-10), 137.3 (C-11), 127.0 (C-11a), 120.3 (C-11b), 64.3 (C-13), 44.0 (C-14); HRFABMS (3-nitrobenzyl alcohol) m/z: 294.1226 [M + H]+ (calcd for C17H16N3O 294.1243); FABMS (3-nitrobenzyl alcohol) (rel. int.) m/z: 294 [M + H] + ; Indonesian ascidian Eusynstyela latericius31

Purple solid; UV (MeOH-TFA) kmax (log e): 222 (sh, 4.43), 244 (4.28), 278 (4.55), 294 (4.60), 372 (sh, 3.57), 388 (3.70), 572 (3.72) nm; UV (MeOH-KOH) kmax (log e): 262 (4.38), 378 (3.97) nm; IR mmax (film): 3072, 1678, 1660, 1619, 1581, 1367, 1199, 1128 cm-1; *H NMR (DMSO-d6, 500 MHz): dH 13.47 (brs, H-1), 8.24 (d, J = 6.5 Hz, H-2), 7.54 (d, J = 6.5 Hz, H-3), 8.20 (d, J = 8.5 Hz, H-4), 7.22 (dd, J = 7.5, 7.5 Hz, H-5), 7.70 (m, H-6), 7.67 (m, H-7), 11.50 (brs, H-8), 7.43 (s, H-10), 10.78 (brs, H-

12), 2.96 (t, J = 6.5 Hz, H-13), 3.24 (dt, J = 7.5, 6.5 Hz, H-14), 8.50 (t, J = 5.5 Hz, H-15), 1.91 (s, H-17); 13C NMR (DMSO-d6, 125 MHz): dC 143.3 (C-2), 104.9 (C-3), 149.2 (C-3a), 113.9 (C-3b), 125.4 (C-4), 122.0 (C-5), 135.0 (C-6), 117.6 (C-7), 141.0 (C-7a), 128.3 (C-8a), 116.2 (C-9), 121.6 (C-10), 136.6 (C-11), 125.9 (C-11a), 120.3 (C-11b), 30.3 (C-13), 37.7 (C-14), 171.4 (C-16), 22.4 (C-17); HRFABMS (3-nitrobenzyl alcohol) m/z: 320.1397 [M + H]+ (calcd for C19H18N3O2 320.1399); FABMS (3-nitrobenzyl alcohol) m/z (rel. int.): 320 [M + H]+ (100), 247 (15); Indonesian ascidian Eusynstyela latericius37

Orange solid; UV (CHCl3) kmax (log e): 248 (4.90), 284 (4.82), 334 (4.22), 350 (4.27), 418 (4.75), 480 (4.41) nm; UV (CHCl3-TFA) kmax (log e): 282 (4.85), 304 (4.89), 422 (4.69), 510 (4.33) nm; IR vmax (film): 3283, 1650, 1630, 1515, 1242 cm-1; *H NMR (DMSO-d6, 500 MHz): dH 8.80 (d, J = 5.5 Hz, H-2), 7.93 (d, J = 5.5 Hz, H-3), 8.33 (d, J = 8.0 Hz, H-4), 7.28 (m, H-5), 7.61 (m, H-6), 7.61 (m, H-7), 12.13 (brs, H-8), 7.38 (brs, H-10), 9.15 (brs, H-12), 9.93 (s, H-13); 13C NMR (DMSO-d6, 125 MHz): dC 151.6 (C-2), 109.3 (C-3), 140.9 (C-3a), 116.7 (C-3b), 124.4 (C-4), 123.0 (C-5), 132.6 (C-6), 117.9 (C-7), 137.1 (C-7a), 134.6 (C-8a), 108.4 (C-9), 113.6 (C-10), 139.0 (C-11), 142.8 (c-11a), 117.9 (C-11b), 191.8 (C-13); HRFABMS (3-nitrobenzyl alcohol) 263.0814 [M + H]+ (calcd for C16H„N2O2 263.0821); FABMS (glycerol) m/z (rel. int.): 263 [M + H]+ (100); Indonesian ascidian Eusynstyela

latericius37

Purple solid; UV (MeOH-TFA) kmax (log e): 222 (4.30), 238 (4.17), 276 (4.37), 294 (4.38), 320 (3.80), 372 (3.54), 386 (3.60), 560 (3.43) nm; UV (MeOH-KOH) kmax (log e): 270 (4.26), 376 (3.95) nm; IR vmax (film): 3072, 1672, 1584, 1202, 1131 cm-1; *H NMR (DMSO-d6, 500 MHz): dH 13.61 (brs, H-1), 8.30 (d, J = 6.5 Hz, H-2), 7.60 (d, J = 6.5 Hz, H-3), 8.23 (d, J = 8.5 Hz, H-4), 7.23 (m, H-5), 7.69 (m, H-6), 7.69 (m, H-7), 10.84 (brs, H-8), 7.47 (s, H-10), 10.98 (brs, H-12), 3.20 (brt, J = 7.0 Hz, H-

13), 3.10 (brm, H-14), 7.95 (brs, H-15); 13C NMR (DMSO-d6, 125 MHz): dC 143.3 (C-2), 105.1 (C-3), 148.9 (C-3a), 113.8 (C-3b), 125.2 (C-4), 122.4 (C-5), 134.7 (C-6), 117.6 (C-7), 141.0 (C-7a), 128.3 (C-8a), 113.8 (C-9), 121.8 (C-10), 137.2 (C-11), 126.7 (C-11a), 120.4 (C-11b), 28.2 (C-13), 37.9 (C-14); HRFABMS (3-nitrobenzyl alcohol) m/z: 278.1280 [M + H]+ (calcd for CnH16N3O 278.1293); FABMS (3-nitrobenzyl alcohol) m/z (rel. int.): 278 [M + H]+ (100), 261 (20), 247 (15); Indonesian ascidian Eusynstyela latericius37

Burnt orange solid; mp 202-204 °C (dec.); UV (MeOH)) kmax (e): 377 (5984), 300 (14,722). 263 (24,874) nm; IR (neat) vmax: 3323, 3066, 2984, 2923, 1651, 1600, 1533 cm-1; *H NMR (CDCl3, 200 MHz): dH 8.46 (dd, J = 8.0, 1.4 Hz, H-1), 7.92 (ddd, J = 8.0, 8.0, 1.2 Hz, H-2), 7.80 (ddd, J = 8.0, 8.0, 1.4 Hz, H-3), 8.27 (dd, J = 8.0, 1.2 Hz, H-4), 8.35 (d, J = 5.6 Hz, H-5), 9.06 (d, J = 5.6 Hz, H-6), 3.73 (brs, H-12), 3.73 (brs, H-13), 6.45 (H-14), 1.91 (H-16), 2.62 (s, SCH3); 13C NMR (DMSO-d6, 50 MHz): dC 122.76 (C-1), 129.63 (C-2), 131.93 (C-3), 118.94 (C-4), 121.28 (C-4a), 136.59 (C-4b), 131.75 (C-5), 149.71 (c-6), 146.39 (c-7a), 179.43 (C-8), 143.39 (C-9), 149.51 (C-10), 151.70 (C-10a), 116.90 (C-10b), 145.49 (C-11a), 30.02 (C-12), 39.75 (C-13), 170.56 (C-15), 23.32 (C-16), 17.86 (SCH3); FABMS m/z: 368 [M + 2H + H] + , 366 [M + H] + ; HRFABMS m/z: 366.1279 [M + H]+ (calcd for C20H19N3O2S); Fiji Islands tunicate Diplosomra sp.29

(continued on next page)

Table 1 (continued)

Isodiplamine (20)

Diplamine B (21)

H3CS^ ^

Nor-segoline (22)

22 COOCH3

Tintamine (23)

(H3C)2N

14 /10 9 17 / S

(H3C)2^^ 13 2

Tintamine diacetate (24)

(H3C)2N

Green solid; mp 208-210 °C; UV-vis (MeOH/TFA) kmax (log e): 215 (4.68), 274 (4.16), 320 (4.01), 406 (4.00) nm; UV-Vis (MeOH/KOH)) kmax (log e): 205 (4.57), 273 (4.20), 320 (4.03), 406 (4.04) nm; IR mmax 3297, 2923, 1652, 1566, 1525, 1430, 1192 cm-1; 1H NMR (DMSO-d6, 300 MHz): dH 8.32 (d, J = 7.8 Hz, H-1), 8.05 (t, J = 7.1 Hz, H-2), 8.02 (obsc, H-3), 9.35 (d, J = 8.1 Hz, H-4), 9.13 (s, H-6), 6.85 (s, H-9), 3.17 (t, J = 6.4 Hz, H-12), 3.52 (obsc, H-13), 7.95 (obsc, H-14), 1.74 (s, H-2'), 2.97 (s, SCH3); 13C NMR (DMSO-d6, 75 MHz): dC 131.7 (C-1), 131.1 (C-2), 129.5 (C-3), 127.5 (C-4), 121.8 (C-4a), 132.8 (C-4b), 138.2 (C-5), 145.9 (C-6), 141.7 (C-7a), 182.6 (C-8), 131.8 (C-9), 151.1 (C-10), 150.1 (c-10a), 129.3 (C-10b), 145.1 (C-11a), 31.2 (C-12), 37.8 (C-13), 169.1 (C-1'), 22.5 (C-2'), 16.4 (SCH3); FABMS m/z: 366 [M + 2H + H] + , 364 [M + H] + ; HRFABMS m/z: 364.1126 [M + H]+ (calcd for C20H18N3O2S 364.1120); New Zealand ascidian Lissoclinum notti36

Dark orange film; UV (MeOH) kmax (log e): 208 (4.39), 265 (3.70), 379 (3.05) nm; IR (neat) mmax: 2924, 2853, 1675, 1202, 1131, 723 cm-1; 1H NMR (cD3OD, 500 MHz): dH 8.34 (d, J = 8.0 Hz, H-1), 7.99 (t, J = 7.7 Hz, H-2), 7.90 (t, J = 7.7 Hz, H-3), 8.80 (d, J = 7.5 Hz, H-4), 8.90 (d, J = 5.0 Hz, H-5), 9.14 (d, J = 4.4 Hz, H-6), 3.89 (t, J = 7.4 Hz, H-12), 3.37 (t, J = 7.2 Hz, H-13), 2.68 (s, H-14); 13C NMR (CD3OD, 125 MHz): dC 132.67 (C-1), 133.32 (C-2), 131.39 (C-3), 124.93 (c-4), 123.10 (C-4a), 139.08 (C-4b), 122.01 (C-5), 150.89 (C-6), 147.71 (C-7a), 181.05 (C-8), 144.73 (C-9), 150.21 (c-10), 150.72 (c-10a), 118.76 (C-10b), 147.07 (C-11a), 29.09 (C-12), 40.17 (C-13), 17.87 (C-14); HRESITOFMS m/z: 322.1005 [M + H] + (calcd for C18H16N3OS 322.1014); Papua New Guinea ascidian Lissoclinum cf. badium38

UV (MeOH) kmax (e): 440 (1200), 400 (2800), 367 (1500), 340 (3000), 264 (12,700), 225 (17,300) nm; UV (MeOH, H+) Xmax (e): 515 (850), 402 (2300), 385 (2300), 366 (1500), 295 (10,900), 264 (12,000), 225 (18,900) nm; IR (CHCl3) vmax: 2940, 1670, 1620, 1520, 1310, 1130, 1070 cm-1; 1H NMR (CDCl3, 360 MHz): dH 8.86 (d, J = 4.8 Hz, H-2), 7.61 (d, J = 4.8 Hz, H-3), 7.18 (dd, J = 7.9, 1.2 Hz, H-4), 7.48 (dt, J = 7.9, 1.2 Hz, H-5), 7.17 (dt, J = 7.9, 1.2 Hz, H-6), 8.03 (d, J = 7.9 Hz, H-7), 11.66 (brs, 8-NH), 7.51 (s, H-10), 4.00 (s, 11-OCH3), 3.99 (s, 12-OCH3); 13C NMR (CDCl3, 90 MHz): dC 151.9 (C-2), 108.7 (C-3), 145.2 (C-3a), 116.5 (C-3b), 130.8 (C-4), 123.9 (C-5), 132.3 (C-6), 122.3 (C-7), 137.9 (c-7a), 137.3 (C-8a), 117.1 (C-9), 109.1 (C-10), 141.1 (C-11), 123.5 (C-11a), 119.3 (C-11b), 168.8 (C-12), 56.2 (11-OCH3), 51.9 (12-OCH3); CIMS (CH4) m/z (re1. int.): 307 [M + H] + (C18H15N2O3, 100); Red Sea tunicate Eudistoma sp.39,40

Yellow powder; IR (KBr) vmax: 3492, 3436, 3270 cm-1; 1H NMR (CDCl3/CD3OD (4:1), 500 MHz): dH 3.75 (t, J = 7.5 Hz, H-2), 2.66 (t, J = 7.5 Hz, H-3), 6.65 (d, J = 1.9 Hz, H-4), 6.72 (dd, J = 8.5, 1.9 Hz, H-6), 8.12 (d, J = 8.5 Hz, H-7), 6.78 (d, J = 1.9 Hz, H-9), 7.18 (brs, H-11), 3.08 (m, H-14), 3.19 (m, H-15), 2.76 (s, H-17); 13C NMR (CDCl3/CD3OD (4:1), 125 MHz): dC 47.4 (C-2), 26.3 (C-3), 136.1 (C-3a), 119.1 (C-3b), 114.3 (C-4), 160.0 (C-5), 113.5 (c-6), 130.6 (C-7), 140.7 (c-7a), 151.6 (C-8a), 111.8 (C-9), 137.2 (C-10), 112.3 (C-11), 167.0 (C-12), 159.6 (C-12a), 142.4 (C-12b), 31.3 (C-14), 59.0 (C-15), 43.1 (C-17); HRFABMS m/z: 368.1390 [M + H]+ (calcd for C20H22N3O2S 368.1432); Indian Ocean tunicate Cystodytes violatinctus41

Amorphous powder; UV (MeOH) kmax (log e): 265 (3.68), 308 (3.67) nm; UV (MeOH + H+) kmax (log e): 280 (3.52), 340 (3.58), 371 (3.65) nm; IR (neat) mmax: 3500, 1762, 1021 cm-1; 1H NMR (CDCl3, 500 MHz): dH 3.95 (t, J = 7.5 Hz, H-2), 2.75 (d, J = 7.5 Hz, H-3), 7.04 (d, J = 1.9 Hz, H-4), 7.09 (dd, J = 8.5, 1.9 Hz, H-6), 8.55 (d, J = 8.5 Hz, H-7), 7.14 (d, J = 1.9 Hz, H-9), 7.72 (d, J = 1.9 Hz, H-11), 3.25 (brt, H-14), 3.38 (brt, H-15), 2.84 (s, H-17), 2.34 (COCH3), 2.43 (COCH3); 13C NMR (CDCl3, 125 MHz): dC 47.7 (C-2), 26.1 (C-3), 119.3 (C-4), 119.4 (C-6), 130.3 (C-7), 119.6 (C-9), 119.7 (C-11), 31.2 (C-14), 59.2 (C-15), 43.3 (C-17), 20.1 (COCH3), 20.8 (COCH3), 169.1 (COCH3); Indian Ocean tunicate Cystodytes violatinctus41

Table 1 (continued)

Varamine A (25)

16\15/ N

10a ,J4b 10b |1 5

Varamine B (26)

^^^ s 6

H,CS^ 7a N

26 OCH3

UV (MeOH) kmax (e): 232 (31,500), 275 (25,800), 292 (sh), 324 (sh), 382 (3530), 464 (51,701), 494 (sh); UV (MeOH, H+) kmax (e): 224 (24,400), 228 (sh), 240 (sh), 263 (sh), 280 (sh), 291 (31,100), 339 (20,400), 359 (4520), 376 (4640), 527 (5670), 552 (sh); IR (CHCl3) vmax: 3450, 3280, 3200, 1650, 1615 cm"1; 'H NMR (CDCl3, 500 MHz): dH 7.87 (dd, J = 8.4, 1.2 Hz, H-1), 7.04 (dd, J = 8.4, 1.3 Hz, H-2), 7.43 (td, J = 8.4, 1.2 Hz, H-3), 7.51 (d, J = 8.4 Hz, H-4), 7.30 (d, J = 5.2 Hz, H-5), 8.67 (d, J = 5.2 Hz, H-6), 10.2 (brs, H-11), 3.30 (m, H-12), 3.30 (m, H-13), 6.22 (brs, H-14), 2.38 (q, J = 6.5 Hz, H-16), 1.29 (t, J = 6.5 Hz, H-17), 4.05 (s, OCH3), 2.53 (s, S-CH3); *H NMR (CD3OD/ TFA-d, 500 MHz): dH 8.09 (d, J = 8.2 Hz, H-1), 7.27 (dd, J = 8.2, 7.3 Hz, H-2), 7.66 (dd, J = 8.3, 7.3 Hz, H-3), 7.77 (d, J = 8.3 Hz, H-4), 7.52 (d, J = 6.5 Hz, H-5), 8.21 (d, J = 6.5 Hz, H-6), 3.23 (m, H-12), 3.33 (m, H-13), 2.35 (q, J = 7.6 Hz, H-16), 1.24 (t, J = 7.6 Hz, H-17), 4.00 (s, OCH3), 2.66 (s, S-CH3); 13C NMR (CD3OD/TFA-d, 125 MHz): dC 126.2 (C-1), 124.8 (C-2), 136.5 (c-3), 119.5 (C-4), 116.0 (C-4a), 132.6 (C-4b), 106.4 (C-5), 143.8 (C-6), 150.5 (C-7a), 140.9 (C-8), 140.5 (C-9), 119.3 (C-10), 134.0 (C-10a), 120.6 (C-10b), 142.2 (C-11a), 29.3 (C-12), 38.8 (C-13), 178.8 (C-15), 30.0 (C-16), 10.4 (C-17), 60.4 (OCH3), 18.7 (S-CH3); FABMS m/z (re1. int.): 394 [M + H] + (100), 293 [M-C5H10NO]+ (43); HRMS m/z: 394.1592 [M + H]+ (calcd for C22H24N3O2S 394.1589); Fiji Island tunicate Lissoclinum vareau9

UV (MeOH) kmax (e): 234 nm (25,900), 274 (21,400), 292 (sh), 324 (sh), 382 (3040), 462 (4320), 491 (sh); UV (MeOH, H+) kmax (e): 223 (20,570), 238 (sh), 266 (sh), 282 (sh), 294 (27,400), 311 (17,300), 362 (sh), 379 (4190), 529 (4720), 552 (4230); IR (CHCl3) vmax: 3450, 3280, 3200, 1650, 1617 cm"1; *H NMR (CD3OD/TFA-d, 500 MHz): dH 8.09 (d, J = 8.2 Hz, H-1), 7.27 (dd, J = 8.2, 7.3 Hz, H-2), 7.66 (dd, J = 8.3, 7.3 Hz, H-3), 7.77 (d, J = 8.3 Hz, H-4), 7.52 (d, J = 6.5 Hz, H-5), 8.21 (d, J = 6.5 Hz, H-6), 3.23 (m, H-12), 3.33 (m, H-13), 2.08 (s, H-16), 4.00 (s, OCH3), 2.66 (s, S-CH3); 13C NMR (CD3OD/TFA-d, 125 MHz): dC 126.2 (C-1), 124.8 (C-2), 136.5 (C-3), 119.5 (C-4), 116.0 (C-4a), 132.6 (C-4b), 106.4 (C-5), 143.8 (C-6), 150.5 (C-7a), 140.9 (C-8), 140.5 (C-9), 119.3 (C-10), 134.0 (C-10a), 120.6 (C-10b), 142.2 (C-11a), 29.3 (C-12), 38.8 (C-13), 175.2 (C-15), 22.5 (C-16), 60.4 (OCH3), 18.7 (S-CH3); FABMS m/z: 380 [M + H]+ (100); HRMS m/z: 380.1420 [M + H]+ (calcd for C21H22N3O2S 380.1433); Fiji Island tunicate Lissoclinum vareau9

25 OCH

Table 2 Pentacyclic pyridoacridine alkaloids.

Kuanoniamine A (27)

Kuanoniamine B (28)

Yellow needles; mp 255-258 °C (dec); UV (MeOH) kmax (log e): 214 (4.26), 224 (4.28), 250

(4.20), 258 (4.19), 295 (3.80), 354 (3.76), 394 (3.61) nm; UV (MeOH, H+) kmax (log e): 208

(4.21), 226 (4.25), 244 (sh, 4.17), 292 (4.17), 382 (3.71) nm; IR (CHCl3) vmax: 3020, 1680, 1590, 1290, 1240, 1220, 1050, 800, 750 cm"1; *H NMR (DMSO-d6, 300 MHz): dH 8.69 (d, J = 6.0 Hz, H-2), 8.45 (d, J = 6.0 Hz, H-3), 8.58 (d, J = 8.1Hz, H-4), 7.60 (dd, J = 8.1, 7.6 Hz, H-5), 7.66 (dd, J = 7.8, 7.6 Hz, H-6), 8.01 (d, J = 7.8 Hz, H-7), 9.28 (s, H-11); 13C NMR (DMSO-d6, 75 MHz): dC 149.04 (C-2), 117.28 (C-3), 137.21 (C-3a), 123.07 (C-3b), 124.08 (C-4), 131.02 (C-5), 132.02 (C-6), 132.02 (C-7), 144.87 (C-7a), 147.27 (C-8a), 176.17 (C-9), 135.21 (C-9a), 162.72 (C-11), 157.84 (C-12a), 147.12 (C-12b), 116.45 (C-12c); HREIMS m/z: 289.0307 (calcd for C16H7N30S 289.0305), 261.0352 (calcd for C15H7N3S 261.0344), 234.0249 (calcd for C14H6N2S 234.0246); EIMS m/z (rel. int.): 261 (42), 234 (14), 190 (16), 117 (13), 69 (18), 58 (100); Micronesian tunicate and its predator, a prosobranch mollusk Chelynotus semperi28

Yellow amorphous powder; mp >300 °C; UV (MeOH) kmax (log e): 204 (4.24), 240 (4.32), 264 (4.18), 294 (sh, 3.89), 344 (3.87), 360 (3.87), 450 (3.44) nm; UV (MeOH+) kmax (log e): 204 (4.17), 238 (4.21), 270 (4.08), 306 (4.27), 344 (3.65), 360 (3.73), 530 (3.41) nm; IR (CHCl3) mmax: 3620, 1640, 1450, 1220, 1110, 1040 cm"1; 1H NMR (DMSO-d6, 300 MHz): dH 8.69 (d, J = 5.0 Hz, H-2), 7.72 (d, J = 5.0 Hz, H-3), 8.09 (d, J = 7.8 Hz, H-4), 7.05 (ddd, J = 7.8, 7.2, 0.6 Hz, H-5), 7.51 (dd, J =8.1, 7.2 Hz, H-6), 7.45 (dd, J = 8.1, 0.6 Hz, H-7), 10.25 (s, H-8), 9.09 (s, H-11), 3.10 (t, J = 7.2 Hz, H-13), 3.34 (td, J = 7.2, 5.2 Hz, H-14), 8.40 (t, J = 5.2 Hz, H-15), 2.00 (m, H-17), 2.00 (m, H-18), 0.83 (d, J = 7.0 Hz, H-19, 20); 13C NMR (DMSO-d6, 75 MHz): dC 150.70 (C-2), 108.26 (C-3), 139.13 (C-3a), 115.61 (C-3b), 123.66 (C-4), 120.80 (C-5), 131.57 (C-6), 116.14 (C-7), 139.17 (c-7a), 133.39 (C-8a), 104.37 (C-9), 139.70 (C-9a), 148.50 (C-11), 140.59 (C-12a), 143.42 (c-12b), 117.58 (C-12c), 30.94 (C-13), 36.39 (C-14), 172.91 (C-16), 44.44 (C-17), 25.17 (C-18), 22.01 (C-19, 20); HREIMS m/z: 402.1520 (calcd for C23H22N4OS 402.1526); EIMS m/z (rel. int.): 402 (19), 314 (20), 301 (21), 289 (72), 261 (78), 234 (23), 190 (22); Micronesian tunicate and its predator, a prosobranch mollusk Chelynotus semperi28

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Table 2 (continued)

Dehydrokuanoniamine B (29)

Kuanoniamine C (30)

Kuanoniamine D (31)

N-Deacetylkuanoniamine C (32)

Orange solid; UV (MeOH) Xmax (e): 237 (50,967), 261 (30 1201,344 (16,413), 359 (16,953), 452 (6047) nm; UV (MeOH, TFA) kmax (e): 277 (43,640), 267 (20,720), 306 (40,333), 359 (11,447), 527 (5940) nm; IR (film) vmax: 3284, 3186, 3151, 3129, 3071, 3058, 3036, 3026, 2921, 2851, 1641, 1632, 1567, 1452 cm"1; 'H NMR (DMSO-d6-TFA-d, 500 MHz): dH 8.46 (d, J = 6.5 Hz, H-2), 7.87 (d, J = 6.5 Hz, H-3), 8.27 (d, J = 8.2 Hz, H-4), 7.27 (ddd, J = 8.2, 5.3, 2.1 Hz, H-5), 7.72 (m, H-6), 7.72 (m, H-7), 11.71 (brs, H-8), 9.43 (s, H-11), 3.12 (t, J = 7.3 Hz, H-13), 3.31 (td, J = 7.3, 5.3 Hz, H-14), 8.35 (t, J = 5.3 Hz, H-15), 5.63 (septet, J = 1.1 Hz, H-17), 2.15 (d, J = 1.1 Hz, H-19), 1.79 (d, J = 1.1 Hz, H-20); 13C NMR (DMSO-d6-TFA-d, 125 MHz): dC 142.72 (C-2), 107.57 (C-3), 148.50 (C-3a), 114.13 (C-3b), 125.53 (C-4), 122.98 (C-5), 135.30 (C-6), 117.5 (C-7), 140.48 (C-7a), 132.80 (C-8a), 108.44 (C-9), 143.15 (C-9a), 153.83 (C-11), 132.74 (C-12a), 131.80 (C-12b), 118.25 (c-12c), 30.98 (C-13), 36.19 (C-14), 167.84 (C-16), 118.08 (C-17), 150.30 (C-18), 19.50 (C-19), 26.84 (C-20); FABMS (glycerol) m/z: 401 [M + H] + , 302, 288; HRFABMS (glycerol) m/z: 401.1429 [M + H]+ (calcd for C23H21N4OS 401.1436); Fijian ascidian Cystodytes sp.35

Yellow amorphous powder; mp >300 °C; UV (MeOH) kmax (log e): 206 (4.24), 240 (4.32), 264 (4.20), 294 (sh, 3.90), 344 (3.88), 359 (3.86), 450 (3.30) nm; UV (MeOH, H+) kmax (log e): 206 (4.16), 240 (4.22), 270 (4.07), 306 (4.27), 360 (3.75), 526 (3.40) nm; IR (CHCl3) mmax: 3580, 3150, 1645, 1590, 1370, 1310, 1140 cm"1; *H NMR (DMSO-d6, 300 MHz): dH 8.68 (d, J = 5.0 Hz, H-2), 7.71 (d, J = 5.0 Hz, H-3), 8.08 (d, J = 7.7 Hz, H-4), 7.05 (dt, J = 7.7, 4.0 Hz, H-5), 7.43 (d, J = 4.0 Hz, H-6), 7.43 (d, J = 4.0 Hz, H-7), 10.10 (s, H-8), 9.07 (s, H-11), 3.08 (t, J = 7.2 Hz, H-13), 3.30 (td, J = 7.2, 5.2, H-14), 8.27 (t, J = 5.2 Hz, H-15), 2.06 (q, J = 7.2 Hz, H-17), 0.96 (t, J = 7.2 Hz, H-18); 13C NMR (DMSO-d6, 100 MHz,): dC 143.0 (C-2), 108.2 (c-3), 139.4 (C-3a), 114.3 (C-3b), 125.4 (C-4), 122.8 (C-5), 135.0 (C-6), 117.6 (C-7), 140.4 (C-7a), 133.5 (C-8a), 107.7 (C-9), 140.2 (c-9a), 153.2 (C-11), 140.7 (C-12a), 143.5 (C-12b), 118.3 (C-12c), 36.3 (C-13), 30.9 (C-14), 174.6 (C-15), 28.4 (C-16), 9.7 (C-17); HREIMS m/z: 374.1270 (calcd for C21H18N4OS 374.1339), 289.0311 (calcd for C16H7N3OS 289.0313); EIMS m/z (re. int.): 374 (5%), 370 (10), 313 (15), 299 (46), 289 (52), 278 (47), 261 (61), 234 (11), 190 (13), 169 (12), 149 (49), 86 (77), 69 (100); Micronesian tunicate and its predator, a prosobranch mollusk Chelynotus semperi and Indonesian sponge Oceanapia sp.21,23,42

Yellow amorphous powder; mp >300 °C; UV (MeOH) kmax (log e): 206 (4.24), 240 (4.32), 264 (4.20), 344 (3.88), 358 (3.87), 452 (3.38) nm; UV (MeOH, H+) kmax (log e): 206 (4.15), 240 (4.22), 270 (4.07), 306 (4.27), 360 (3.75), 526 (3.39); IR (CHCl3) vmax: 3610, 3000, 2940, 2840, 1640, 1590, 1460, 1240, 1210, 1100, 1060, 1000 cm"1; 1H NMR (DMSO-d6, 300 MHz): dH 8.70 (d, J = 5.1 Hz, H-2), 7.73 (d, J = 5.1 Hz, H-3), 8.11 (d, J = 8.1 Hz, H-4), 7.06 (dt, J = 8.1, 3.9 Hz, H-5), 7.45 (d, J = 3.9 Hz, H-6), 7.45 (d, J = 3.9 Hz, H-7), 10.15 (s, H-8), 9.09 (s, H-11), 3.07 (t, J = 7.2 Hz, H-13), 3.27 (td, J = 7.2, 5.2, H-14), 8.39 (t, J = 5.2 Hz, H-15), 1.87 (s, H-17); 13C NMR (DMSO-d6, 75 MHz): dC 151.01 (C-2), 108.57 (C-3), 139.33 (C-3a), 115.78 (C-3b), 124.01 (C-4), 121.07 (C-5), 131.92 (C-6), 116.34 (C-7), 139.33 (C-7a), 133.49 (C-8a), 104.56 (C-9), 139.59 (C-9a), 149.01 (C-11), 139.86 (C-12a), 143.56 (c-12b), 117.74 (C-12c), 31.08 (C-13), 36.74 (c-14), 170.96 (C-16), 22.52 (C-17); HREIMS m/z: 360.1007 (calcd for C20H16N4OS 360.1045), 288.0590 (calcd for C17H10N3S 288.0595); EIMS m/z: 360 (40), 356 (18), 314 (26), 301 (31), 299 (30), 288 (100); Micronesian tunicate and its predator, a prosobranch mollusk Chelynotus semperi28

Orange amorphous powder; UV (MeOH) kmax (log e): 201 (4.14), 239 (4.33), 264 (4.11), 295 (4.08), 304 (4.05), 342 (3.82), 355 (3.85) nm; 1H NMR (DMSO, 300 MHz): dH 8.67 (d, J = 5.1 Hz, H-2), 7.68 (d, J = 5.1 Hz, H-3), 8.06 (d, J = 7.9 Hz, H-4), 7.02 (m, H-5), 7.40 (m, H-6, 7), 9.08 (s, H-11), 3.08 (br, H-13), 2.95 (br, H-14); 13C NMR (DMSO, 400 MHz): dC 150.9 (C-2), 108.4 (C-3), 139.4 (C-3a), 115.8 (C-3b), 123.9 (C-4), 120.8 (C-5), 131.8 (C-6), 116.3 (C-7), 139.6 (C-7a), 133.8 (C-8a), 106.0 (C-9), 140.0 (C-9a), 148.9 (C-11), 140.7 (c-12a), 143.5 (C-12b), 117.9 (C-12c), 34.6 (C-13), 39.6 (C-14); ESIMS m/z: 319 [M + H] + ; ESIMS/MS m/z: 302 [319-NH3] + ; HREIMS m/z: 318.0930 (calcd for C18H14N4S 318.0939); The Micronesian sponge Oceanapia sp.23

Table 2 (continued)

Kuanoniamine F (33)

Dehydrokuanoniamine F (34)

Sagitol (35)

// Il I 12c HO

Sagitol C (36)

Yellow gum; [a]D +105 (c 0.18, CH3OH); UV (MeOH) kmax (log e): 206 (4.26), 242 (4.49), 262 (4.29), 296 (sh, 3.98), 346 (4.04), 363 (4.05), 455 (3.38) nm; IR (KBr) vmax: 3271, 2966, 1647, 1573, 1469, 1413, 1260, 831, 758 cm"1; 1H NMR (CDCl3, 500 MHz): dH 8.81 (d, J = 4.7 Hz, H-2), 7.50 (d, J = 4.7 Hz, H-3), 7.93 (d, J = 8.0 Hz, H-4), 7.05 (t, J = 7.5 Hz, H-5), 7.42 (t, J = 7.5 Hz, H-6), 7.50 (brd, J = 6.6 Hz, H-7), 10.20 (brs, H-8), 8.77 (s, H-11), 3.13 (dd, J = 6.8, 5.2 Hz, H-13), 3.42 (dd, J = 6.8, 5.2 Hz, H-14), 6.22 (brs, H-15), 2.26 (dd, J = 13.8, 6.9 Hz, H-17), 1.24 (d, J = 6.9 Hz, H-18), 1.78 (dd, J = 13.8, 7.4 Hz, H-19), 1.56 (dd, J = 13.8, 7.4 Hz, H-19), 0.98 (t, J = 7.4 Hz, H-20); 1H NMR (DMSO-d6, 500 MHz): dH 8.70 (d, J = 4.8 Hz, H-2), 7.72 (d, J = 4.8 Hz, H-3), 8.10 (d, J = 7.9 Hz, H-4), 7.06 (brt, J = 5.2 Hz, H-5), 7.46 (brt, J = 5.2 Hz, H-6), 7.49 (brt, J = 5.2 Hz, H-7), 10.24 (brs, H-8), 9.11 (s, H-11), 3.36 (t, J = 7.2 Hz, H-14), 3.09 (brt, J = 7.2 Hz, H-13), 8.38 (brs, H-15), 2.16 (ddd, J = 13.8, 6.9 Hz, H-17), 0.99 (d, J = 6.9 Hz, H-18), 1.52 (dd, J = 7.4, 6.9 Hz, H-19), 1.31 (dd, J = 7.4, 6.9 Hz, H-19), 0.76 (t, J = 7.4 Hz, H-20); 13C NMR (DMSO-d6, 125 MHz): dH 150.6 (C-2), 108.2 (C-3), 139.3 (C-3a), 115.7 (C-3b), 123.7 (C-4), 120.7 (C-5), 131.5 (C-6), 116.0 (C-7), 139.7 (C-7a), 133.3 (c-8a), 104.5 (C-9), 139.3 (C-9a), 148.6 (C-11), 140.5 (C-12a), 143.4 (C-12b), 117.6 (C-12c), 30.9 (C-13), 36.2 (C-14), 176.8 (C-16), 41.1 (C-17), 17.1 (C-18), 26.5 (C-19), 11.5 (c-20); ( + )-ESIMS m/z: 403 [M + H] + , 401 [M-H] + ; ( + )-HRESIMS m/z: 403.1598 (calcd for C23H23N4OS 403.1593); Singaporean ascidian43

Purple oil; UV (MeOH, H+) kmax (log e): 229 (4.0), 268 (3.9), 307 (4.2), 317.7 (4.3), 358 (4.3), 528 (3.2) nm; IR (RBr) vmax: 3310, 3030, 2921, 1653, 1424 cm"1; 1H NMR (DMSO-d6, 400 MHz): dH 7.85 (m, H-1), 8.55 (m, H-2), 11.24 (brs, H-7), 7.68 (m, H-8), 7.68 (m, H-9), 7.23 (m, H-10), 8.26 (d, J = 8.4 Hz, H-11), 3.15 (t, J = 7.1 Hz, H-12), 3.39 (H-13), 8.30 (t, J = 5.1 Hz, H-14), 6.38 (q, J = 6.2 Hz, H-17), 1.68 (d, J = 6.2 Hz, H-18), 1.72 (s, H-19), 9.35 (s, H-2'); 13C NMR (DMSO-d6, 100 MHz): dC 107.9 (C-1), 144.8 (C-2), 135.3 (C-4), 144.0 (C-5), 107.4 (C-6), 133.0 (C-6a), 140.3 (C-7a), 117.3 (c-8), 134.4 (c-9), 122.5 (c-10), 127.1 (C-11), 114.6 (C-11a), 152.4 (C-11b), 118.2 (c-11c), 31.0 (C-12), 36.8 (C-13), 169.6 (C-15), 131.0 (C-16), 130.7 (C-17), 13.7 (C-18), 12.4 (C-19), 151.0 (C-2'); ESIMS m/ z: 401 [M + H] + ; HRESIMS m/z: 401.1448 [M + H]+ (calcd for C23H20N4OS, 401.1436); South-Pacific Ocean ascidian Cystodytes violatinctus42

Orange residue; UV (MeOH) kmax: 264, 340 nm; 1H NMR (DMSO-d6, 400 MHz): dH 8.87 (d, J = 5.8 Hz, H-2), 8.49 (d, J = 5.8 Hz, H-3), 8.72 (d, J = 7.8 Hz, H-4), 7.81 (t, J = 7.5 Hz, H-5), 7.97 (t, J = 7.8 Hz, H-6), 8.27 (d, J = 7.8 Hz, H-7), 9.29 (s, H-11), 2.90 (m, H-13A), 2.49 (m, H-13B), 2.93 (m, H-14A), 2.75 (m, H-14B), 1.71 (q, J = 7.3 Hz, H-16), 0.79 (t, J = 6.9 Hz, H-17); 13C NMR (DMSO-d6, 100 MHz): dC 150.5 (C-2), 116.2 (C-3), 140.1 (C-3a), 122.3 (C-3b), 123.5 (C-4), 128.5 (C-5), 132.5 (C-6), 129.8 (C-7), 146.5 (c-7a), 162.5 (C-8a), 72.8 (c-9), 146.1 (C-9a), 155.6 (c-11), 151.5 (C-12b), 115.1 (C-12c), 46.5 (C-13), 35.4 (C-14), 174.2 (C-15), 28.2 (C-16), 9.5 (C-17); ESIMS m/z: 391 [M + H] + ; Indonesian sponge Oceanapia sp.21

Orange residue; UV (MeOH) kmax: 254, 366 nm; 1H NMR (DMSO-d6, 400 MHz): dH 9.15 (d, J = 5.4 Hz, H-2), 8.92 (d, J = 5.4 Hz, H-3), 9.05 (dd, J = 8.2, 1.3 Hz, H-4), 8.06 (dt, J = 8.2, 1.3 Hz, H-5), 8.11 (dt, J = 8.2, 1.3 Hz, H-6), 8.47 (dd, J = 8.2, 1.3 Hz, H-7), 9.72 (s, H-11), 3.20 (m, H-13), 2.78 (m, H-14), 2.61 (m, H-16), 1.25 (d, J = 6.9 Hz, H-17), 1.25 (d, J = 6.9 Hz, H-18); 13C NMR (DMSO-d6, 100 MHz): dC 149.1 (C-2), 117.2 (C-3), 140.5 (C-3a), 122.5 (C-3b), 124.0 (C-4), 131.6 (C-5), 131.8 (C-6), 132.0 (C-7), 146.5 (C-7a), 162.7 (C-8a), 72.5 (C-9), 146.3 (C-9a), 162.1 (C-11), 149.0 (C-12a), 152.0 (C-12b), 115.5 (C-12c), 47.2 (C-13), 41.5 (C-14), 174.3 (C-15), 39.9 (C-16), 26.3 (c-17), 26.3 (C-18); HRESIMS m/z 405.1464 [M + H] + (calcd for C22H21N4O2S, 405.1463); Indonesian

sponge Oceanapia sp.2

(continued on next page)

Table 2 (continued)

Petrosamine (37)

Debromopetrosamine trifluoroacetate salt (38)

Ecionine A (39)

Ecionine B (40)

Dark blue needles; mp 330 °C; UV (MeOH) kmax (e): 286 (36,418), 346 (11,732), 369 (11,183), 414 (6161) nm; IR (film) vmax: 3056, 1644, 1583, 1531, 1494, 1422, 924, 838 cm"1; 1HNMR (DMSO-d6, 300 MHz): dH 8.37 (d, J = 2.0 Hz, H-1), 7.92 (dd, J = 9.2, 2.0 Hz, H-3), 9.16 (d, J = 9.2 Hz, H-4), 4.75 (s, H-6), 9.88 (s, H-9), 9.17 (d, J = 5.3 Hz, H-11), 9.29 (d, J = 5.3 Hz, H-12), 4.60 (s, H-14), 3.83 (s, H-15, 16); 13C NMR (DMSO-d6, 75 MHz): dC 131.8 (C-1), 122.9 (C-2), 135.1 (C-3), 126.3 (C-4), 120.0 (C-4a), 128.8 (C-4b), 187.2 (C-5), 69.8 (C-6), 114.2 (C-7a), 161.3 (C-8), 139.9 (C-8a), 145.6 (C-9), 142.5 (C-11), 121.7 (C-12), 131.5 (C-12a), 141.4 (C-12b), 114.3 (C-12c), 142.1 (C-13a), 48.3 (C-14), 53.1 (C-15, 16); HRFABMS m/z: 422.0505 [M + H] + (calcd for C21H17O2N3Br 422.0504); Thai marine sponge Petrosia n. sp.44

Purple-blue amorphous solid; UV (MeOH) kmax (log e): 216 (4.49), 282 (4.74), 374 (3.38), 592 (3.15) nm; IR (film) vmax: 3063, 1682, 1648, 1588, 1498, 1206, 1129, 801, 722 cm"1; 1H NMR (DMSO-d6, 500 MHz): dH 8.19 (d, J = 9.0 Hz, H-1), 7.73 (dd, J = 9.0, 7.5 Hz, H-2), 7.84 (dd, J = 8.0, 7.5 Hz, H-3), 9.27 (d, J = 8.0 Hz, H-4), 4.41 (s, H-6), 9.72 (s, H-9), 8.75 (d, J = 6.5 Hz, H-11), 9.39 (d, J = 6.5 Hz, H-12), 4.53 (s, H-14), 3.84 (s, H-15), 3.84 (s, H-16); 13C NMR (DMSO-d6, 125 MHz): dC 143.0 (C-1), 128.4 (C-2), 133.5 (C-3), 125.1 (c-4), 125.2 (C-4a), 115.8 (C-4b), 187.2 (C-5), 71.4 (C-6), 115.1 (C-7a), 161.0 (C-8), 132.7 (C-8a), 145.9 (C-9), 142.0 (C-11), 122.9 (C-12), 143.5 (C-12a), 139.5 (C-12b), 129.9 (C-12c), 143.1 (C-13a), 49.0 (C-14), 54.1 (C-15), 54.1 (C-16); HRESIMS m/z 344.1415 [M + H]+ (calcd for C2iHigN3O2 344.1399); Sponge Xestospongia cf. carbonaria45

Light brown solid; UV (MeOH) kmax (log e): 215 (4.23), 228 (4.26), 284 (4.00), 319 (3.70), 376 (3.80), 443 (3.28) nm; IR (KBr) vmax: 1666, 1462, 1201, 1136 cm"1; 1H NMR (DMSO-d6, 500 MHz): dH 8.41 (d, J = 7.8 Hz, H-1), 8.17 (dd, J = 7.8, 7.2 Hz, H-2), 8.09 (dd, J = 7.8, 7.2 Hz, H-3), 9.07 (d, J = 7.8 Hz, H-4), 9.16 (d, J = 5.4 Hz, H-5), 9.28 (d, J = 5.4 Hz, H-6), 10.83 (s, H-7), 11.12 (s, 8-NH), 2.92 (t, J = 7.8 Hz, H-10), 4.11 (dd, J = 7.8, 7.2 Hz, H-11), 11.57 (brs, H-12); 13C NMR (DMSO-d6, 125 MHz): dC 131.1 (C-1), 132.9 (C-2), 131.6 (C-3), 124.7 (C-4), 122.8 (C-4a), 136.8 (C-4b), 121.3 (C-5), 149.5 (c-6), 142.5 (C-7a), 157.7 (C-8), 99.0 (C-8a), 193.2 (C-9), 34.6 (C-10), 40.5 (C-11) 157.9 (c-12a), 142.0 (C-12b), 116.2 (C-12c), 144.1 (C-13a); ( + )-LRESIMS m/z (rel. int.): 301 [M"CF3COO"]+ (100); ( + )-HRESIMS m/z: 301.1079 [M-CF3COO"]+ (calcd for C18H13N4O 301.1084); Australian marine sponge Ecionemia geodides16

Light brown solid; UV (MeOH) kmax (log e): 211 (3.66), 235 (3.74), 286 (3.48), 321 (3.20), 382 (3.20), 447 (3.00); UV (MeOH + KOH) kmax (log e): 212 (3.97), 241 (3.73), 293 (3.40), 366 (3.08), 431 (2.97), 541 (2.77) nm; IR (KBr) mmax: 3410, 1674, 1547, 1432, 1203, 1135, 1025, 994 cm"1; 1H NMR (DMSO-d6, 500 MHz): dH 10.61 (s, 1-OH), 7.51 (d, J = 7.8 Hz, H-2), 7.96 (dd, J = 8.4, 7.8 Hz, H-3), 8.46 (d, J = 8.4 Hz, H-4), 9.08 (d, J = 5.4 Hz, H-5), 9.26 (d, J = 5.4 Hz, H-6), 10.86 (s, H-7), 11.10 (s, 8-NH), 2.95 (t, J = 7.8 Hz, H-10), 4.20 (dt, J = 7.8, 7.8 Hz, H-11), 11.61 (s, H-12); 13C NMR (DMSO-d6, 125 MHz): dC 155.8 (c-1), 115.1 (C-2), 133.2 (C-3), 114.6 (C-4), 123.6 (C-4a), 136.9 (C-4b), 121.6 (C-5), 149.4 (C-6), 142.6 (C-7a), 157.6 (C-8), 98.9 (C-8a), 193.4 (C-9), 34.5 (c-10), 39.9 (C-11), 157.7 (c-12a), 139.2 (C-12b), 116.4 (C-12c), 133.5 (C-13a); ( + )-LRESIMS m/z (rel. int.): 317 [M-CF3COO"]+ (100); ( + )-HRESIMS m/z: 317.1039 [M"CF3COO"]+ (calcd for C18H13N4O2 317.1033); Australian marine sponge Ecionemia geodides16

Table 2 (continued)

Cystodimine A (41)

Cystodimine B (42)

Shermilamine A (43)

Debromoshermilamine A = Shermilamine B (44)

Yellow solid; UV (MeOH) kmax (log e): 228 (4.2), 282 (3.8), 318 (3.7), 384 (3.8), 440 (3.3) nm; *H NMR (CD3OD, 400 MHz): dH 9.22 (d, J = 5.5 Hz, H-2), 8.98 (d, J = 5.5 Hz, H-3), 8.93 (dd, J = 8.1, 1.1 Hz, H-4), 8.03 (td, J = 8.1, 1.1 Hz, H-5), 8.09 (td, J = 8.1, 1.1 Hz, H-6), 8.44 (dd, J = 8.0, 1.7 Hz, H-7), 2.90 (t, J = 7.3 Hz, H-11), 4.15 (t, J = 7.3 Hz, H-12), 11.81 (13-NH), 10.66 (brs, 14a-NH), 11.04 (brs, 14b-NH); 13C NMR (CD3OD, 100 MHz): dC 150.8 (C-2), 122.2 (C-3), 138.8 (C-3a), 124.6 (C-3b), 125.3 (C-4), 133.1 (C-5), 133.9 (C-6), 130.0 (C-7), 146.2 (C-7a), 143.9 (C-8a), 159.0 (C-9), 100.4 (C-9a),

194.6 (C-10), 35.7 (C-11), 41.9 (C-12), 160.7 (C-13a), 143.5 (C-13b), 117.6 (C-13c); ( + )-ESIMS m/z: 301 [M + H] + ; HRFABMS m/z: 301.1088 (calcd for C18H13N4O, 301.1089); Western Mediterranean ascidian Cystodytes dellechiajei24

Yellow solid; UV (MeOH) kmax (log e): 228 (3.8), 278 (3.5), 308 (3.5), 388 (3.1), 450 (3.4) nm; *H NMR (CD3OD, 400 MHz): dH 9.17 (d, J = 5.6 Hz, H-2), 8.82 (d, J = 5.5 Hz, H-3), 8.10 (d, J = 2.5 Hz, H-4), 7.60 (dd, J = 8.7, 2.5 Hz, H-6), 8.31 (d, J = 8.7 Hz, H-7), 3.00 (t, J = 7.3 Hz, H-11), 4.16 (t, J = 7.3 Hz, H-12); 13C NMR (CD3OD, 100 MHz): dC 150.1 (C-2), 122.2 (C-3), 137.8 (C-3a), 127.0 (C-3b), 108.5 (C-4), 163.2 (C-5), 124.3 (C-6), 135.3 (c-7), 141.0 (C-7a), 139.5 (C-8a), 159.4 (C-9), 132.5 (C-9a), 194.9 (C-10), 35.7 (C-11), 41.8 (C-12), 160.7 (C-13a), 143.2 (C-13b), 117.5 (C-13c); ( + )-ESIMS m/z: 317 [M + H] + ; HRFABMS m/z: 317.1028 (calcd for C18H13N4O2 317.1039); Western Mediterranean ascidian Cystodytes dellechiajei24

Orange prisms; mp >300 °C; UV (MeOH) kmax (log e): 238 (4.52), 282 (4.45), 298 (4.39), 350 (3.90), 392 (3.71), 470 (3.76) nm; UV (MeOH, H+) kmax (log e): 242 (4.40), 286 (4.36), 312 (4.55), 320 (4.54), 364 (3.74), 382 (3.75), 536 (3.77) nm; IR (CHCl3) vmax: 3680, 1665, 1650, 1630, 1605, 1595, 1500, 1460, 1435, 1375, 1335, 1010, 930, 830 cm"1; *H NMR (DMSO-d6, 300 MHz): dH 8.43 (d, J = 4.8 Hz, H-2), 7.39 (d, J = 4.8 Hz, H-3), 7.86 (d, J = 7.8 Hz, H-4), 7.07 (dd, J = 8.7, 1.8 Hz, H-5), 7.49 (d, J = 1.8 Hz, H-7), 10.17 (s, H-8), 3.54 (s, H-11), 9.22 (s, H-13), 2.80 (m, H-14), 3.05 (m, H-15), 8.55 (t, J = 4.8 Hz, H-16), 1.90 (s, H-18); 13C NMR (DMSO-d6, 75 MHz): dC 150.57 (C-2), 107.46 (C-3), 138.49 (C-3a), 114.71 (C-3b), 125.90 (C-4), 123.33 (C-5), 124.97 (C-6), 118.22 (C-7), 141.10 (C-7a), 130.44 (C-8a), 109.32 (C-9), 122.01 (C-9a), 29.26 (C-11), 163.48 (C-12), 121.31 (C-13a), 136.55 (C-13b), 116.27 (C-13c), 27.62 (C-14), 37.03 (C-15), 171.53 (C-17), 22.45 (C-18); HREIMS m/z: 468.0260 (calcd for C21Hl779BrN4O2S 468.0255); EIMS m/z (rel. int.): 470 (85), 468 (82), 398 (55), 396 (65), 207 (60), 177 (100), 147 (30), 135 (40), 91 (25), 73 (30), 57 (68); Tunicate Trididemnum sp.46

Mp >310 °C; UV (MeOH) kmax (e): 460 (3700), 390 (4100), 365 (6000), 284 (16,700), 270 (20,000), 264 (27,800) nm; UV (MeOH+) kmax (e): 535 (2000), 385 (4100), 320 (20,000), 300 (27,000), 283 (27,000), 268 (27,000) nm; IR (CHCl3) mmax: 3920, 2860, 1660, 1630, 1440, 1350 cm"1; *H NMR (DMSO-d6, 360 MHz): dH 8.51 (H-2), 7.40 (H-3), 8.06 (H-4), 7.45 (H-5), 7.03 (H-6), 7.35 (H-7), 10.2 (s, H-8), 3.57 (s, H-11), 9.20 (s, H-13), 2.97 (m, H-14), 3.11 (m, H-15), 8.61 (H-16), 1.93 (s, H-18); 13C NMR (DMSO-d6, 90 MHz): dC 151.0 (C-2), 107.3 (C-3), 139.7 (c-3a), 115.5 (C-3b), 124.2 (C-4), 121.1 (C-5), 132.1 (C-6), 116.5 (c-7), 140.2 (C-7a), 131.3 (C-8a), 116.7 (C-8b), 108.9 (C-9), 121.7 (C-9a), 29.3 (C-11),

163.7 (C-12), 121.6 (C-13a), 137.0 (C-13b), 27.3 (C-14), 37.1 (C-15), 171.7 (C-17), 22.3 (C-18); CIMS (methane) m/z (rel. int.): 391 [M + H]+ (C21Hl9N4O2S, 100); Tunicate Eudistoma sp.40

Fine orange prisms; mp 254 °C; UV (MeOH) kmax (log e): 234 (4.54), 282 (4.45), 298

(4.39), 348 (3.97), 390 (3.76), 468 (3.81) nm; UV (MeOH, H+) kmax (log e): 232 (4.44), 282

(4.40), 302 (4.53), 318 (4.69), 364 (3.78), 382 (3.79), 536 (3.83) nm; IR (CHCl3) vmax: 3290, 3265, 2980, 1640, 1630, 1600, 1580, 1550, 1450, 1420, 1350, 1310, 1190, 1120, 1070, 900, 840, 800, 740, 650 cm"1; 1H NMR (DMSO-d6, 300 MHz): dH 8.50 (d, J =5.1 Hz, H-2), 7.49 (d, J =5.1 Hz, H-3), 8.01 (dd, J = 7.8, 1.2 Hz, H-4), 7.02 (ddd, J = 7.8, 7.8, 1.2 Hz, H-5), 7.44 (ddd, J = 7.8, 7.8, 1.2 Hz, H-6), 7.44 (ddd, J = 7.8, 7.8, 1.2 Hz, H-7), 10.24 (s, H-8), 3.33 (s, H-11), 9.26 (s, H-13), 2.82 (m, H-14), 3.00 (m, H-15), 8.57 (t, J = 4.8 Hz, H-16), 1.92 (s, H-18); 13C NMR (DMSO-d6, 75 MHz): dC 150.56 (C-2), 107.7 (C-3), 139.3 (C-3a), 115.34 (C-3b), 123.87 (C-4), 120.76 (C-5), 131.80 (C-6), 116.43 (C-7), 139.90 (C-7a), 131.07 (C-8a), 108.56 (C-9), 121.40 (C-9a), 29.25 (C-11), 163.41 (C-12), 121.21 (C-13a), 136.60 (C-13b), 116.27 (C-13c), 27.66 (C-14), 37.07 (C-15), 171.62 (C-17), 22.38 (C-18); HREIMS m/z: 390.1204 (calcd for C21Hl8N4O2S 390.1259), 318.0645 (calcd for C18H12N3OS 318.0590); EIMS m/z (rel. int.): 390 (100), 318 (80), 212 (45), 162 (90); Tunicate Trididemnum sp.46

(continued on next page)

Table 2 (continued)

N-Deacetylshermilamine B (45)

N^ 12^. O

Shermilamine C (46)

Shermilamine D (47)

Purple solid; UV (MeOH) kmax (log e): 231 (4.5), 279 (4.2), 298 (4.8), 314 (4.9), 358 (3.6), 378 (3.6), 540 (3.9) nm; 1H NMR (DMSO-d6, 400 MHz): dH 8.55 (d, J = 5.3 Hz, H-2), 7.63 (d, J = 5.3 Hz, H-3), 8.14 (d, J = 8.1 Hz, H-4), 7.13 (t, J = 7.9 Hz, H-5), 7.53 (t, J = 7.9 Hz, H-6), 7.48 (d, J = 8.1 Hz, H-7), 9.81 (brs, 8-NH), 3.64 (s, H-11), 9.55 (s, 13-NH), 3.24 (t, J = 8.1 Hz, H-14), 2.98 (bt, H-15), 7.98 (bs, H-16); 13C NMR (DMSO-d6, 100 MHz): dC 150.5 (C-2), 107.3 (C-3), 142.5 (C-3a), 115.4 (C-3b), 125.0 (C-4), 122.2 (C-5), 132.7 (C-6), 116.9 (C-7), 140.0 (C-7a), 131.5 (C-8a), 108.4 (C-9), 124.4 (c-9a), 29.4 (c-11), 164.0 (C-12), 121.7 (C-13a), 136.0 (C-13b), 117.1 (C-13c), 25.8 (C-14), 36.9 (C-15); ( + )-ESIMS m/z: 349 [M + H] + ; ( + )-ESIMS/MS (from the precursor ion m/z 349) 332 [M + H"NH3] + ; HRFABMS m/z: 349.1140 (calcd for C19H17N4OS 349.1123); Western Mediterranean ascidian Cystodytes dellechiajei24

Orange solid; UV (MeOH) kmax (e): 231 (9185), 279 (6669), 296 (5693), 350 (2683), 390 (1896), 463 (1664); UV (MeOH, TFA) kmax (e): 218 (12,414), 277 (6033), 300 (7491), 318 (8123), 359 (17,031, 382 (1716), 538 (1423); IR (film) 3285, 3203, 3052, 2925, 2854, 1634, 1433 cm"'; 1H NMR (DMSO-d6-TFA-d, 500 MHz): dH 8.35 (d, J = 5.9 Hz, H-2), 7.45 (d, J = 5.9 Hz, H-3), 8.04 (d, J = 7.9 Hz, H-4), 7.11 (dd, J = 7.9, 7.9 Hz, H-5), 7.55 (dd, J = 7.9, 7.9 Hz, H-6), 7.51 (d, J = 7.9 Hz, H-7), 11.37 (brs, H-8), 3.58 (s, H-11), 9.64 (brs, H-13), 2.89 (t, J = 7.2 Hz, H-14), 3.08 (m, H-15), 8.56 (t, J = 5.7 Hz, H-16), 5.70 (t, J = 1.1 Hz, H-18), 2.22 (s, H-20), 1.84 (s, H-21); 13C NMR (DMSO-d6-TFA-d, 125 MHz): dC 146.06 (C-2), 105.92 (C-3), 144.48 (C-3a), 114.74 (C-3b), 124.78 (C-4), 122.22 (C-5), 133.78 (C-6), 116.97 (C-7), 140.09 (C-7a), 131.45 (C-8a), 111.46 (C-9), 127.97 (C-9a), 29.49 (C-11), 164.27 (C-12), 117.86 (C-13a), 130.56 (C-13b), 117.14 (C-13c), 27.97 (c-14), 36.61 (c-15), 168.31 (C-17), 117.92 (C-18), 150.81 (C-19), 19.60 (C-20), 26.95 (C-21); FABMS (glycerol) m/z: 431 [M + H] + , 332, 318; HRFABMS (glycerol) m/z: 431.1529 [M + H]+ (calcd for C24H23N4O2S 431.1542); Fijian ascidian Cystodytes sp.35

Foaming oil; 1H NMR (CDCl3, 400 MHz): dH 8.40 (d, J = 4.8 Hz, H-2), 7.20 (d, J = 4.7 Hz, H-3), 7.80 (d, J = 7.9 Hz, H-4), 6.98 (t, J = 7.5 Hz, H-5), 7.33 (brt, J = 7.5 Hz, H-6), 6.84 (d, J = 8.1 Hz, H-7), 3.50 (s, H-11), 2.96 (brt, J = 4.0 Hz, H-14), 2.62 (brt, J = 4.8 Hz, H-15), 2.50 (s, H-17), 2.50 (s, H-18), 10.20 (brs, 8-NH), 9.05 (brs, 13-NH); 13C NMR (CDCl3, 100 MHz): dC 150.7 (C-2), 106.8 (C-3), 140.2 (C-3a), 116.3 (C-3b), 123.7 (C-4), 120.6 (C-5), 131.7 (C-6), 116.4 (c-7), 140.7 (C-7a), 132.9 (C-8a), 116.7 (c-8b), 109.6 (C-9), 121.4 (C-9a), 29.7 (C-11), 163.5 (C-12), 121.3 (C-13a), 137.3 (C-13b), 27.4 (C-14), 58.7 (C-15), 44.9 (C-17), 44.9 (C-18); Tunicate Cystodytes violatinctus47

Shermilamine E (48)

8* J91

NO(Me)2

Shermilamine F (49)

N^ rJO

Brown amorphous powder; UV (MeOH) kmax (log e): 232 (4.29), 296 (4.10), 349 (3.70), 391 (3.51), 464 (3.49) nm; UV (MeOH + H+) kmax (log e): 230 (4.25), 281 (4.18), 300 (4.21), 315 (4.25), 361 (3.53), 382 (3.50), 526 (3.45) nm; IR (neat) vmax: 3500, 3326, 1623 cm"1; 1H NMR (CDCl3/CD3OD (4:1), 500 MHz): dH 8.44 (d, J = 5.0 Hz, H-2), 7.20 (d, J = 5.0 Hz, H-3), 7.78 (brd, J = 8.0 Hz, H-4), 6.99 (brt, J = 7.5 Hz, H-5), 7.34 (brt, J = 7.5 Hz, H-6), 7.10 (brd, J = 8.2 Hz, H-7), 3.50 (brs, H-11), 3.40 (brt, H-14), 3.42 (brt, H-15), 3.29 (s, H-17), 11.80 (brs, 8-NH), 9.10 (brs, 13-NH); 13C NMR (CDCl3/ CD3OD (4:1), 125 MHz): dC 150.8 (C-2), 107.1 (C-3), 140.1 (C-3a), 115.9 (C-3b), 123.5 (C-4), 121.1 (C-5), 131.9 (C-6), 116.8 (C-7), 140.4 (C-7a), 132.2 (C-8a), 116.9 (C-8b), 106.0 (c-9), 120.7 (C-9a), 29.9 (C-11), 163.6 (C-12), 121.2 (C-13a), 137.7 (c-13b), 22.3 (C-14),

68.2 (C-15), 58.5 (C-17); HRFAMS m/z: 393.1402 [M-

Cystodytes violatinctus41

H] ; Indian Ocean tunicate

Purple oil; UV (MeOH, H+) kmax (log e): 230 (4.2), 278 (3.9), 301 (4.1), 317.7 (4.3), 361 (3.4), 381 (3.4), 524 (3.3) nm; IR (RBr) vmax: 3297, 3061, 2922, 1642, 1536, 1479 cm"1; 1H NMR (CDCl3, 400 MHz): dH 7.29 (d, J = 6.2 Hz, H-1), 8.41 (d, J = 6.2 Hz, H-2), 11.87 (brs, H-7), 7.85 (d, J = 8.3 Hz, H-8), 7.66 (dd, J = 8.1, 8.3 Hz, H-9), 7.26 (dd, J = 8.1, 8.3 Hz, H-10), 7.96 (d, J = 8.1 Hz, H-11), 3.21 (m, H-12), 3.37 (m, H-13), 6.70 (t, J = 7.2 Hz, H-14), 6.78 (q, J = 7.0 Hz, H-17), 1.87 (d, J = 7.0 Hz, H-18), 1.97 (s, H-19), 3.45 (s, H-2'), 10.33 (brs, H-4'); 13C NMR (CDCl3, 100 MHz): dC 104.9 (C-1), 144.6 (C-2), 129.3 (C-3a), 118.2 (C-4), 130.1 (C-5), 111.7 (C-6), 132.2 (C-6a), 140.8 (C-7a), 118.5 (c-8), 134.6 (c-9), 123.0 (C-10), 124.3 (C-11), 115.0 (C-11a), 147.0 (C-11b), 118.5 (C-11c), 28.3 (C-12), 38.0 (C-13), 171.5 (C-15), 130.1 (C-16), 133.7 (c-17), 14.2 (C-18), 12.2 (C-19), 30.2 (c-2'), 164.1 (C-3'); ESIMS m/z: 431 [M + H] + ; HRESIMS m/z: 431.1529 [M + H] + (calcd for C24H22N4O2S 431.1542); South-Pacific Ocean ascidian Cystodytes violatinctus43

Table 2 (continued)

Sebastianine A (50)

Lissoclinidine trifluoroacetate salt (51)

CF3CO2H ♦

Lissoclinidine B (52)

/ Tl ^li™,

Labuanine A (53)

Ascididemin (54)

13a N 13

Yellow amorphous solid; UV (MeOH) kmax (log e): 223 (4.56), 238 (4.49), 265 (4.36), 301 (4.11), 390 (3.85) nm; IR (neat) vmax: 1683, 1590, 1208, 1139, 844, 803, 724 cm"1; 1H NMR (DMSO-d6, 400 MHz): dH 8.18 (d, J = 7.5 Hz, H-1), 7.94 (t, J = 7.1 Hz, H-2), 7.81 (t, J =7.1 Hz, H-3), 8.88 (d, J = 7.6 Hz, H-4), 8.96 (d, J = 5.5 Hz, H-5), 9.19 (d, J = 5.5 Hz, H-6), 12.85 (s, H-9), 7.48 (d, J = 2.6 Hz, H-10), 7.01 (d, J = 2.6 Hz, H-11); 13C NMR (DMSO-d6, 100 MHz): dC 129.8 (C-1), 131.6 (C-2), 128.0 (C-3), 124.0 (C-4), 121.3 (C-4a), 137.0 (C-4b), 119.7 (C-5), 149.1 (C-6), 148.5 (C-7a), 171.9 (C-8), 129.5 (C-8a), 128.9 (C-10), 106.3 (C-11), 130.9 (C-11a), 148.5 (C-11b), 117.1 (C-11c), 145.2 (C-12a);

EIMS m/z: 271.0752 [M] +

dellechiajei48

(calcd for Ci7H9N3Q 271.0745); Brazilian ascidian Cystodytes

Hygroscopic purple solid; UV-vis (MeOH/TFA) Xmax (log e): 218 (4.41), 281 (4.26), 295 (4.21), 310 (4.10), 384 (3.61), 548 (3.40) nm; UV-Vis (MeOH/KOH)) kmax (log e): 225 (4.17), 264 (4.16), 301 (4.00), 312 (3.99), 374 (3.23), 391 (3.38), 486 (3.32) nm; IR mmax: 3442, 2913, 1660, 1590, 1423, 1200, 1129 cm"1; 1H NMR (DMSO-d6, 400 MHz): dH 7.75 (m, H-1), 7.75 (m, H-2), 7.31 (m, H-3), 8.24 (d, J =8.2 Hz, H-4), 7.44 (d, J = 6.7 Hz, H-5), 8.18 (d, J = 6.6 Hz, H-6), 13.2 (brs, H-7), 11.71 (brs, H-11), 2.93 (t, J = 7.3 Hz, H-12), 3.22 (td, J = 7.3, 5.5 Hz, H-13), 8.47 (t, J = 5.5 Hz, H-14), 1.91 (s, H-2'), 6.09 (s, OCH2S); 13C NMR (DMSO-d6, 100 MHz): dC 117.8 (C-1), 135.0 (C-2), 123.2 (C-3), 125.3 (C-4), 114.2 (C-4a), 148.3 (C-4b), 103.6 (C-5), 143.2 (C-6), 121.1 (C-7a), 134.2 (c-8), 137.3 (c-9) 109.7 (C-10), 131.8 (C-10a), 118.3 (C-10b), 140.1 (C-11a), 30.1 (C-12), 36.3 (C-13), 171.3 (C-1'), 22.4 (C-2'), 77.2 (OCH2S); FABMS m/z: 364 [M + H] + ; HRFABMS m/z: 364.1122 (calcd for C20H18N3O2S 364.1120); New Zealand ascidian Lissoclinum notti36

Purple film; UV (MeOH) kmax (log e): 214 (4.37), 265 (3.98), 298 (3.81), 309 (3.78), 389 (3.15), 477 (3.10) nm; IR (neat) vmax: 2923, 2852, 1681, 1204, 1135, 723 cm"1; 1H NMR (cD3OD, 500 MHz): dH 7.82 (d, J = 8.4 Hz, H-1), 7.76 (t, J = 7.5 Hz, H-2), 7.38 (t, J = 7.5 Hz, H-3), 8.20 (d, J = 8.4 Hz, H-4), 7.45 (d, J = 6.6 Hz, H-5), 8.13 (d, J = 6.6 Hz, H-6), 3.29 (m, H-12), 3.24 (m, H-13), 6.11 (s, H-14); 13C NMR (CD3OD, 125 MHz,): dC 119.27 (C-1), 136.45 (C-2), 125.04 (C-3), 126.16 (C-4), 116.19 (C-4a), 150.76 (C-4b), 105.08 (C-5), 143.95 (C-6), 124.67 (C-7a), 136.60 (C-8), 139.78 (C-9), 108.43 (C-10), 133.79 (C-10a), 120.40 (C-10b), 141.86 (C-11a), 29.30 (C-12), 38.07 (C-13), 78.88 (c-14); HRESTOFMS m/z: 322.1003 ([M + H]+ (calcd for C18H16N3OS 322.1014); Papua New Guinea ascidian Lissoclinum cf. badium38

Yellow amorphous solid; UV (MeOH) kmax (e): 357 (6067), 317 (5190), 288 (7720), 227 (18,000) nm; IR (KBr) vmax: 3410, 2360, 1682, 1210 cm"1; 1H NMR (DMSO-d6, 500 MHz): dH 8.41 (d, J = 7.9 Hz, H-1), 8.18 (dd, J = 7.9, 7.3 Hz, H-2), 8.11 (dd, J = 7.9, 7.3 Hz, H-3), 9.09 (d, J = 7.9 Hz, H-4), 9.18 (d, J = 5.5 Hz, H-5), 9.30 (d, J = 5.5 Hz, H-6), 2.93 (t, J = 7.6 Hz, H-10), 4.11 (t, J = 7.6 Hz, H-11); 13C NMR (DMSO-d6, 125 MHz,): dC 131.0 (C-1), 132.9 (C-2), 131.5 (C-3), 124.5 (C-4), 122.8 (C-4a), 136.8 (C-4b), 121.2 (C-5), 149.4 (C-6), 142.6-142.4 (C-7a), 157.8-157.6 (C-8), 99.0 (c-8a), 193.5 (c-9), 34.5 (C-10), 40.5 (C-11), 157.8-157.6 (C-12a), 142.6-142.4 (c-12b), 116.2 (C-12c), 144.0 (C-13a); FABMS m/z: 302 [M + H] + ; HRFABMS m/z: 302.0904 (calcd for C18H12N3O2 302.0929); Indonesian marine sponge Biemna fortis26

Yellow solid; mp >300 °C; UV (MeOH)) kmax (e): 220 (49,500), 248 (48,000), 273 (sh, 27,500), 298 (17,000), 308 (15,700), 340 (sh, 11,300), 377 (13,600) nm; IR (KBr) vmax: 1680, 1600, 1580, 1410, 1260, 860, 740 cm"1; 1H NMR (CDCl3/CD3OD, 500 MHz): dH 8.55 (dd, J = 7.7, 1.3 Hz, H-1), 8.05 (ddd, J = 8.1, 7.7, 1.3 Hz, H-2), 7.99 (ddd, J = 8.1, 7.7, 1.3 Hz, H-3), 8.76 (dd, J = 7.7, 1.3 Hz, H-4), 8.69 (d, J = 5.6 Hz, H-5), 9.22 (d, J = 5.6 Hz, H-6), 9.14 (dd, J = 4.7, 1.7 Hz, H-9), 7.75 (dd, J = 7.7, 4.7 Hz, H-10), 8.79 (dd, J = 7.7, 1.7 Hz, H-11); 13C NMR (CDCl3/CD3OD, 125 MHz): dC 132.7 (C-1), 132.52 (C-2), 131.53 (C-3), 123.59 (C-4), 123.91 (C-4a), 138.51 (C-4b), 117.70 (c-5), 149.87 (c-6), 149.87 (c-7a), 152.38 (C-7b), 155.67 (C-9), 126.30 (C-10), 136.99 (C-11), 129.24 (c-11a), 181.99 (C-12), 145.94 (C-12a), 118.24 (C-12b), 145.75 (C-13a); HRFABMS m/z: 286.1006 [M + 2H + H] + ; EIMS m/z: 283 [M] + , 255 [M-CO] + , 228 [M"CO"HCN] + , 200 [M—CO—2HCNI+ [34]; Okinawan tunicate Didemnum sp.48 and Western Mediterranean ascidian Cystodytes dellechiajei49

(continued on next page)

O „ OH

Table 2 (continued)

3-Hydroxyascididemin (55)

8,9-Dihydro-11-hydroxyascididemin (56)

11-Hydroxyascididemin (57)

57 O OH

12-Deoxyascididemin (58)

9-Hydroxyisoascididemin (59)

'H NMR (CD3OD/CDCI3 (1:1), 600 MHz): dH 8.37 (d, J = 9.0 Hz, H-1), 7.55 (dd, J = 9.0, 2.4 Hz, H-2), 8.02 (d, J = 2.4 Hz, H-4), 8.56 (d, J = 5.7 Hz, H-5), 9.14 (d, J = 5.7 Hz, H-6), 9.09 (dd, J = 4.8, 1.8 Hz, H-9), 7.73 (dd, J = 7.8, 4.8 Hz, H-10), 8.77 (dd, J = 7.8, 1.8 Hz, H-11); 13C NMR (CD3OD/CDCl3 (1:1), 150 MHz): dC 134.99 (C-1), 123.93 (C-2), 161.69 (C-3), 107.24 (C-4), 126.86 (C-4a), 137.79 (C-4b), 118.44 (C-5), 149.06 (c-6) 149.58 (C-7a), 152.57 (C-7b), 155.61 (C-9), 126.62 (C-10), 137.24 (C-11), 129.86 (C-11a), 182.29 (C-12), 142.96 (C-12a), 118.90 (C-12b), 141.03 (C-13a); ( + )-EIMS m/z: 299.0 [M] + ; Calliactis parasitica (Actiniaria)50

Yellow amorphous powder; mp >300 °C; UV (MeOH) kmax (e): 218 (17,000), 273 (9300), 319 (6700), 355 (5300) nm; IR (RBr) vmax: 3250, 2900, 1660, 1600, 1020 cm"1; *H NMR (CDC13/CD3OD (1:1), 400 MHz): dH 8.16 (dd, J = 8.0, 1.5 Hz, H-1), 7.81 (ddd, J = 8.0, 7.4, 1.1 Hz, H-2), 7.74 (ddd, J = 8.0, 7.4, 1.5 Hz, H-3), 8.50 (dd, J = 8.0, 1.1 Hz, H-4), 8.41 (d, J = 5.7 Hz, H-5), 8.94 (d, J = 5.7 Hz, H-6), 3.82 (t, J = 7.8 Hz, H-9), 2.59 (t, J = 7.8 Hz, H-10); 13C NMR (CDC13/CD3OD (1:1), 100 MHz): dC 132.0 (C-1), 132.5 (c-2), 131.5 (C-3), 124.5 (C-4), 119.0 (C-5), 150.5 (C-6), 41.0 (C-9), 37.0 (C-10); EIMS m/z (rel. int.): 301 [M]+ (85), 271 (20), 243 (8); HRFABMSm/z: 302.0928 [M + H]+ (calcdfor C18Hl2N3O2 302.0930); Okinawan marine sponge Biemna sp.51

Yellow amorphous solid; mp >250 °C; UV (MeOH) kmax (e): 203 (25,000), 227 (38,000), 275 (18,000), 285 (17,000), 370 (11,000); IR (KBr) vmax: 3067, 1674 cm"1; 1H NMR (CDCl3, 300 MHz): dH 8.64 (dd, J = 8.1, 1.8 Hz, H-1), 8.06 (dt, J = 8.1, 1.2 Hz, H-2), 8.00 (dt, J = 8.1, 1.8 Hz, H-3), 9.31 (d, J = 8.1, 1.2 Hz, H-4), 8.58 (d, J = 5.7 Hz, H-5), 9.31 (d, J = 5.7 Hz, H-6), 7.15 (d, J = 5.7 Hz, H-9), 8.89 (d, J = 5.7 Hz, H-10), 13.06 (s, OH); 13C NMR (CDCl3, 75 MHz): dC 133.3 (C-1), 132.1 (c-2), 131.5 (C-3), 123.1 (C-4), 137.8 (C-4b), 117.2 (C-5), 150.1 (C-6), 149.3 (C-7a), 154.1 (C-8), 156.7 (C-9), 114.8 (C-10), 169.5 (c-11), 115.6 (c-11a), 181.8 (C-12), 117.7 (C-12b); LRMS m/z (rel. int.): 299.0 [M] + (100), 271.0 [M-H2O]+ (47); HRFABMS m/z: 300.0768 [M + H] + ; Ascidian Amphicarpa meridiana and Leptoclinides sp.13

1H NMR (CDCl3/CD3OD (1:1), 400 MHz): dH 8.64 (dd, J = 8.1, 1.8 Hz, H-1), 8.06 (dt, J = 8.1, 1.2 Hz, H-2), 8.00 (dt, J = 8.1, 1.8 Hz, H-3), 8.73 (dd, J = 8.1, 1.2 Hz, H-4), 8.58 (d, J = 5.7 Hz, H-5), 9.31 (d, J = 5.7 Hz, H-6), 7.15 (d, J = 5.7 Hz, H-9), 8.89 (5, J = 5.7 Hz, H-10); 13C NMR (CDCl3/CD3OD (1:1), 100 MHz): dC 133.3 (C-1), 132.1 (C-2), 131.5 (C-3), 123.1 (C-4), 117.2 (C-5), 150.1 (C-6), 156.7 (C-9), 114.8 (C-10); Okinawan marine sponge Biemna sp.51

Brown amorphous solid; UV (MeOH) kmax (log e): 219 (2.83), 239 (2.80), 271 (sh, 2.72), 299 (sh, 2.50), 342 (2.38), 400 (2.10) nm; IR (KBr) vmax: 3684, 2924, 2854, 2360, 2341, 1383, 669, 651 cm"1; 1H NMR (DMSO-d6, 400 MHz): dH 7.43 (d, J =8.4 Hz, H-1), 7.68 (dd, J =8.4, 7.2 Hz, H-2), 7.22 (dd, J = 7.8, 7.2 Hz, H-3), 8.34 (d, J = 7.8 Hz, H-4), 8.23 (d, J = 6.0 Hz, H-5), 8.75 (d, J = 6.0 Hz, H-6), 8.90 (dd, J = 4.2, 1.2 Hz, H-9), 7.84 (dd, J = 8.4, 4.2 Hz, H-10), 8.55 (dd, J = 8.4, 1.2 Hz, H-11), 7.35 (s, H-12), 12.35 (s, H-13); 13C NMR (DMSO-d6, 100 MHz): dC 116.6 (C-1), 134.9 (C-2), 121.9 (C-3), 125.5 (C-4), 114.3 (C-4a), 147.2 (C-4b), 111.3 (C-5), 143.9 (C-6), 137.9 (C-7a), 130.6 (C-7b), 145.4 (C-9), 125.8 (C-10), 135.8 (C-11), 131.1 (C-11a), 99.4 (C-12), 135.4 (C-12a), 120.2 (C-12b), 140.0 (C-13a); ( + )-LRESIMS m/z: 270 [M + H] + ; ( + )-HRESIMS m/z: 270.1030 [M + H]+ (calcd for C18H12N3 270.1026); Australian ascidian Polysyncraton echinatum22

Yellow amorphous solid; UV (MeOH) kmax (e): 365 (5270), 277 (10,600), 227 (15,000) nm; 1H NMR (CDCl3, 500 MHz): dH 8.61 (d, J = 7.9 Hz, H-1), 7.96 (dd, J = 7.9, 7.3 Hz, H-2), 7.85 (dd, J = 7.9, 7.3 Hz, H-3), 8.62 (d, J = 7.9 Hz, H-4), 8.70 (d, J = 5.5 Hz, H-5), 9.31 (d, J = 5.5 Hz, H-6), 7.13 (d, J = 5.5 Hz, H-10), 8.91 (d, J = 5.5 Hz, H-11); 1H NMR (CF3COOD, 500 MHz): dH 9.53 (d, J = 6.1 Hz), 9.35 (d, J = 6.1 Hz), 8.91 (d, J = 7.9 Hz), 8.89 (d, J = 7.0Hz), 8.54 (d, J = 7.9 Hz), 8.29 (t, J = 7.9 Hz), 8.18 (t, J = 7.9 Hz), 7.64 (d, J = 7.0 Hz); 13C NMR (CF3COOD, 125 MHz): dC 179.0, 175.0,

147.3, 147.2, 146.6, 145.2, 141.8, 139.4, 137.8, 137.3, 135.4, 133.9, 126.9, 126.0, 121.4,

118.4, 117.7, 114.2; FABMS m/z: 322 [M + Na] + ; HRFABMS m/z: 322.0587 (calcd for C18H9N3O2Na 322.0593); Indonesian marine sponge Biemna fortis26

Table 2 {continued)

11r^ 10

Yellow amorphous solid; 'H NMR (DMSO-d6, 900 MHz): dH 8.36 (d, J = 7.8 Hz, H-1), 8.04 (t, J = 7.8 Hz, H-2), 7.92 (t, J = 7.8 Hz, H-3), 8.93 (d, J = 7.8 Hz, H-4), 8.95 (d, J = 5.4 Hz, H-5), 9.21 (d, J = 5.4 Hz, H-6), 6.64 (d, J = 7.2 Hz, H-10), 8.00 (d, J = 7.2 Hz, H-11); 13C NMR (DMSO-d6, 225 MHz): dC 130.9 (C-1), 132.0 (C-2), 129.5 (C-3), 124.2 (C-4), 122.2 (C-4a), 136.7 (C-4b), 119.2 (C-5), 150.0 (C-6), 147.6 (C-7a), 179.9 (c-8) 120.4 (C-8a), 176.0 (C-9), 122.4 (C-10), 131.7 (C-11), 150.2 (C-12a), 147.6 (C-12b), 117.2 (C-12c), 144.8 (C-13a); *H NMR (CDCl3, 600 MHz): dH 8.61 (d, J = 7.9 Hz, H-1), 7.96 (dd, J = 7.9, 7.3 Hz, H-2), 7.85 (dd, J = 7.9, 7.3 Hz, H-3), 8.62 (d, J = 7.9 Hz, H-4), 8.70 (d, J = 5.5 Hz, H-5), 9.31 (d, J = 5.5 Hz, H-6), 7.13 (d, J = 5.5 Hz, H-10), 8.91 (d, J = 5.5 Hz, H-11); ( + )-HRESIMS m/z: 322.0587 [M + Na]+ (calcd for C18H9N3O2Na 322.0591); Australian marine sponge Ancorina geodides52

Meridine (6D)

O 8 TI 4ЬТ

60 iN„

Meridin-12(13H)-one (6l)

61 7 N

Yellow amorphous solid; mp >250 °C; FT IR {KBr) vmax: 3444, 3071, 1692, 1605 cm"1; 'h NMR {CDCl3, 300 MHz): dH 7.25 {d, J = 5.5 Hz, H-11), 8.79 {d, J = 5.5 Hz, H-10), 8.23 {d, J =8.1 Hz, H-1), 7.97 {dd, J = 8.1, 7.8 Hz, H-2), 7.86 {dd, J = 8.3, 7.8 Hz, H-3), 8.64 {d, J = 8.3 Hz, H-4), 8.66 {d, J = 5.6 Hz, H-5), 9.38 {d, J = 5.6 Hz, H-6), 15.26 {s, OH); 13C NMR {CDCl3, 75 MHz): de 132.6 {C-1), 129.6 {C-2), 129.4 {C-3), 123.3 {C-4), 121.7 {C-4a), 137.9 {C-4b), 119.7 {c-5), 151.5 {C-6), 147.6 {C-7a), 180.3 {C-8), 148.9 {C-8a), 153.7 {C-10), 116.8 {C-11), 167.3 {C-12), 116.4 {C-l2a), 152.1 {C-l2b), 117.9 {C-l2c), 142.6 {C-l3a); LRMS m/z {rel. int.): 299.1 [M]+ {100), 271.0 [M-CO]+ {l0.4); South Australia marine ascidian Amphicarpa meridiana and Leptoclinides sp.13

'H NMR (CDCl3, 300 MHz): dH 8.10 (d, J =8.1 Hz, H-11), 6.53 (d, J =8.1 Hz, H-10), 7.44 (d, J = 8.1 Hz, H-1), 7.63 (t, J = 8.1 Hz, H-2), 7.37 (d, J = 8.1 Hz, H-3), 8.20 (d, J =8.1 Hz, H-4), 7.64 (d, J = 5.4 Hz, H-5), 8.94 (d, J = 5.4 Hz, H-6), 12.51 (s, NH); South Australia marine ascidian Amphicarpa meridiana and Leptoclinides sp.13

9-Aminobenzo[b]pyrido[4,3,2-de][1,10]phenanthrolin-8(8H)-one (62)

Yellow amorphous solid; UV (MeOH) kmax (e): 373 (7020), 282 (14,100), 231 (20,000) nm; *H NMR (DMSO-d6, 500 MHz): dH 8.35 (d, J = 7.9 Hz, H-1), 8.01 (dd, J = 7.9, 7.3 Hz, H-2), 7.90 (dd, J = 7.9, 7.3 Hz, H-3), 8.94 (d, J = 7.9 Hz, H-4), 9.03 (d, J = 5.5 Hz, H-5), 9.25 (d, J = 5.5 Hz, H-6), 6.95 (d, J = 6.1 Hz, H-10), 8.42 (d, J = 6.1 Hz, H-11), 7.95 (br, NH2), 8.94 (br, NH2); FABMS m/z: 299 [M + H] + ; HRFABMS m/z: 299.0951 (calcd for CigHnN4O 299.0933); Indonesian marine sponge Biemna fortis26

Ancorine A (63) HO. "

Dark-yellow amorphous solid; UV (MeOH) kmax (log e): 234 (4.14), 282 (3.88), 378 (3.60) nm; 1H NMR (DMSO-d6, 900 MHz): dH 7.45 (d, J = 8.1 Hz, H-2), 7.85 (t, J = 8.1 Hz, H-3), 8.39 (d, J =8.1 Hz, H-4), 8.94 (d, J = 5.4 Hz, H-5), 9.24 (d, J = 5.4 Hz, H-6), 6.49 (d, J = 7.2 Hz, H-10), 8.02 (t, J = 7.2 Hz, H-11), 12.67 (brs, H-12), 10.47 (s, 1-OH); 13C NMR (DMSO-d6, 225 MHz): dC 155.2 (C-1), 115.5 (C-2), 131.8 (C-3), 114.3 (C-4), 123.5 (C-4a), 136.9 (C-4b), 119.8 (C-5), 150.4 (C-6), 146.9 (C-7a), 178.6 (C-8), 119.4 (C-8a), 175.7 (C-9), 122.4 (C-10), 136.7 (C-11), 146.8 (C-12a), 142.0 (C-12b), 116.9 (C-12c), 133.3 (C-13a); HRESIMS m/z: 338.0541 [M + Na]+ (calcd for C18H9N3O3Na 338.0536); Australian marine sponge Ancorina geodides52

(continued on next page)

б DMSO-rf

Table 2 (continued)

Cnemidine A (64)

Arnoamine A (65)

10 11 65

Arnoamine B (66)

Arnoamine C (67)

Arnoamine D (68)

Yellow amorphous solid; UV (MeOH) kmax (log e): 230 (4.10), 276 (3.77), 374 (3.53) nm; *H NMR (DMSO-d6, 600 MHz): dH 8.43 (d, J = 7.5 Hz, H-1), 8.09 (t, J = 7.5 Hz, H-2), 8.01 (t, J = 8.0 Hz, H-3), 9.01 (d, J = 8.1 Hz, H-4), 8.98 (d, J = 5.5 Hz, H-5), 9.21 (d, J = 5.5 Hz, H-6), 7.99 (d, J = 8.5 Hz, H-9), 6.59 (d, J = 8.5 Hz, H-10); 13C NMR (DMSO-d6, 150 MHz): dC 131.6 (C-1), 132.1 (C-2), 130.3 (C-3), 124.0 (C-4), 122.5 (C-4a), 136.8 (C-4b), 118.1 (C-5), 147.1 (c-6), 146.7 (C-7a), 149.6 (C-7b), 146.6 (C-9), 121.7 (C-10), 174.3 (C-11), 122.2 (C-11a), 178.9 (C-12), 145.6 (C-12a), 116.9 (C-12b), 145.1 (C-13a), 48.9 (8-CH3); HRESIMS m/z: 336.0742 [M + Na]+ (calcd for C19H„N3O2Na 336.0743); Australian tunicate Cnemidocarpa stolonifera52

Yellow glass; IR (film) vmax: 3418 (broad), 3094, 1681, 1200 cm"1; UV (0.5% TFA in MeOH) Xmax (log e): 278 (4.11), 401 (3.09), 486 (3.17) nm; (in 50 mM KOH in MeOH) kmax (log e): 258 (4.15), 328 (3.89), 431 (3.05) nm; *H NMR (TFA-d/CDCl3): dH 8.28 (d, J = 8.4 Hz, H-1), 8.12 (t, J = 7.6 Hz, H-2), 7.80 (t, J = 7.8 Hz, H-3), 8.68 (d, J = 7.8 Hz, H-4), 8.35 (d, J = 6.1 Hz, H-5), 8.84 (d, J = 6.1 Hz, H-6), 8.20 (brs, H-9), 7.51 (d, J = 3.0 Hz, H-10), 8.49 (d, J = 3.0 Hz, H-10); 13C NMR (TFA-d/CDCl3): dC 116.3 (C-1), 135.7 (C-2), 126.1 (C-3), 127.1 (C-4), 117.9 (C-4a), 141.7 (C-4b), 109.6 (C-5), 137.5 (c-6), 126.3 (C-7a), 113.4 (C-7b), 139.8 (C-8), 115.6 (C-9), 119.7 (C-9a), 119.5 (C-9b), 110.7 (C-10), 121.3 (C-11), 135.8 (C-11b); EIMS m/z (rel. int.): 258 [M]+ (100), 229 (67), 201 (10), 129 (36); HREIMS m/z: 258.0794 (calcd for C17H10N2O 258.0791); Ascidian Cystodytes sp.53

Yellow glass; IR (film) vmax: 3094, 1681, 1200 cm"1; UV (0.5% TFA in MeOH) kmax (log e): 280 (4.15), 398 (3.15), 467 (3.22) nm, (in 50 mM KOH in MeOH) kmax (log e): 254 (4.14), 309 (3.88), 400 (3.17) nm; 1H NMR (TFA-d/CDCl3): dH 8.35 (d, J =8.1 Hz, H-1), 8.15 (t, J = 7.8 Hz, H-2), 7.82 (t, J = 7.5 Hz, H-3), 8.74 (d, J = 7.8 Hz, H-4), 8.47 (brs, H-5), 8.92 (brs, H-6), 8.16 (brs, H-9), 7.60 (brs, H-10), 8.57 (brs, H-11), 4.27 (8-OCH3); 13C NMR (TFA-d/CDCl3): dC 116.4 (C-1), 135.9 (C-2), 126.2 (C-3), 127.4 (C-4), 118.1 (C-4a), 142.0 (c-4b), 110.6 (C-5), 137.8 (C-6), 126.6 (C-7a), 113.5 (C-7b), 143.8 (C-8), 110.4 (C-9), 119.5 (C-9a), 119.3 (C-9b), 110.7 (C-10), 121.3 (C-11), 136.0 (C-11b), 57.0 (8-OCH3); EIMS m/z (rel. int.): 272 [M]+ (83), 271 (100), 257 (19), 242 (66), 229 (52); HREIMS m/z: 272.0946 (calcd for C18H12N2O 272.0947); Ascidian Cystodytes sp.53

Orange oil; UV (MeOH, H+) kmax (log e): 275 (4.0), 278 (4.0), 310 (3.8), 402 (3.4), 473 (3.4) nm; IR (RBr) vmax: 3442, 3232, 2923, 1664, 1559, 1490, 1450 cm"1; 1H NMR (DMSO-d6, 400 MHz): dH 8.63 (d, J = 5.9 Hz, H-1), 9.06 (d, J = 5.9 Hz, H-2), 7.93 (s, H-5), 8.56 (d, J = 7.9 Hz, H-8), 7.97 (dd, J = 7.9, 7.8 Hz, H-9), 7.71 (dd, J = 7.9, 7.8 Hz, H-10), 8.96 (d, J = 7.8 Hz, H-11), 7.22 (s, H-12), 10.51 (s, H-14), 6.83 (q, J = 6.9 Hz, H-17), 1.89 (d, J = 6.9 Hz, H-18), 1.95 (s, H-19); 13C NMR (DMSO-d6, 100 MHz): dC 110.6 (C-1), 141.6 (C-2), 130.0 (C-3a), 143.9 (C-4), 109.1 (C-5), 117.5 (C-6), 113.1 (C-6a), 136.0 (c-7a), 116.6 (C-8), 133.2 (C-9), 124.7 (c-10), 127.0 (C-11), 119.0 (C-11a), 116.4 (C-11c), 106.2 (C-12), 131.4 (C-13), 169.5 (C-15), 131.2 (C-16), 133.0 (C-17), 14.0 (C-18), 12.3 (C-19); ESIMS m/z: 356 [M + H] + ; HRESIMS m/z: 356.1398 [M + H]+ (calcd for C22H18N3O2 356.1399); South-Pacific Ocean ascidian Cystodytes violatinctus43

Orange oil; UV (MeOH, H+) kmax (log e): 275 (4.0), 277 (4.0), 309 (3.8), 403 (3.4), 473 (3.4) nm; IR (RBr) vmax: 3440, 3235, 2922, 2915, 1665, 1559, 1491, 1449 cm"1; 1H NMR (DMSO-d6, 400 MHz): dH 8.74 (d, J = 6.4 Hz, H-1), 9.05 (d, J = 6.4 Hz, H-2), 8.08 (brs, H-5), 8.70 (d, J = 8.5 Hz, H-8), 8.06 (dd, J = 8.5, 8.4 Hz, H-9), 7.76 (dd, J = 8.5, 8.4 Hz, H-10), 9.02 (d, J = 8.4 Hz, H-11), 7.32 (s, H-12), 10.48 (s, H-14), 6.17 (s, H-16), 1.97 (brs, H-18), 2.15 (s, H-19), 10.30 (4-OH); 13C NMR (DMSO-d6, 100 MHz): dC 110.6 (C-1), 140.5 (C-2), 142.9 (C-4), 110.2 (C-5), 117.1 (C-6), 118.0 (C-6a), 136.0 (C-7a), 116.7 (C-8), 133.8 (c-9), 125.0 (C-10), 127.3 (C-11), 118.7 (C-11a), 138.6 (C-11b), 113.0 (C-11c), 106.3 (C-12), 131.4 (C-13), 169.6 (C-15), 117.6 (C-16), 154.5 (C-17), 27.4 (C-18), 19.8 (C-19); ESIMS m/z: 356 [M + H] + ; HRESIMS m/z: 356.1402 [M + H] + ; (calcd for C22H18N3O2 356.1399); South-Pacific Ocean ascidian Cystodytes violatinctus43

DMSO-fl

Table 2 (continued)

Dercitin (69)

13 2 3 3 12 12b 3a 4 11\ i il i i 10 9 7 69 1' N 18 17 Deep violet powder; mp 168 °C; 'H NMR (TFA-d, 360 MHz): dH 8.70 (d, J =7.0 Hz, H-2), 8.17 (d, J = 7.0 Hz, H-3), 8.51 (d, J = 8.3 Hz, H-4), 7.74 (dd, J = 8.3, 6.6 Hz, H-5), 8.10 (dd, J = 8.3, 6.6 Hz, H-6), 7.97 (d, J = 8.3 Hz, H-7), 9.50 (s, H-1l), 5.21 (s, H-13), 4.14 (t, J = 5.7 Hz, H-14), 3.95 (t, J = 5.7 Hz, H-15), 3.56 (s, H-17, 18); 13C NMR (TFA-d, 90 MHz): dC 149.3 (C-2), 109.8 (C-3), 150.9 (C-3a), 116.4 (s, C-3b), 126.5 (C-4), 126.5 (C-5), 138.2 (C-6), 119.1 (C-7), 141.1 (C-7a), 135.5 (C-8a), 107.0 (C-9), 148.2 (C-9a), 151.7 (c-11), 136.1 (C-12a), 136.6 (C-12b), 122.0 (C-12c), 51.4 (C-13), 28.5 (C-14); 56.5 (C-15), 45.2 (C-17, 18); HREIMS m/z (rel. int.): 360.1398 (c21H20N4S) (7), 314.0746 (C19H12N3S) (5), 302.0731 (C18H12N3S) (100), 288.0570 (C17H10N3S) (2); Marine sponge Dercitus sp.27

Nordercitin (70) 2

N^il 3 12 Ji? La 4 X xXX N ia^^^ N ' 7a ^^ 10 ' 13 H 7 70 X 1' 17 Yellow solid; mp 176 °C; *H NMR (TFA-d, 360 MHz): dH 8.58 (H-2), 8.04 (H-3), 8.32 (H-4), 7.56 (H-5), 7.92 (H-6), 7.73 (H-7), 9.58 (H-11), 3.77 (H-13), 3.58 (H-14), 3.14 (H-16, 17); 13C NMR (TFA-d, 90 MHz): dC 143.74 (C-2), 111.05 (C-3), 153.48 (C-3a), 117.24 (C-3b), 127.18 (C-4), 127.78 (C-5), 139.71 (C-6), 120.44 (C-7), 142.21 (C-7a), 137.06 (C-8a), 106.55 (C-9), 145.27 (C-9a), 158.44 (C-11), 130.60 (C-12a), 134.48 (C-12b), 121.00 (c-12c), 32.01 (C-13), 58.49 (C-14), 44.75 (15-NCH3); HRFABMS m/z: 347.1316 [M + H] + (C20H19N4S); Sponges Dercitus sp. and Stetleta sp.18

Dercitamine (71)

N^3 12 12b 3a 4 S -Ji^^A^-OJ^^ 5 / Il j 12c tt ^ ^L A ^ 6 10 I« h 7 71 114 1' NHCH3 15 Orange solid; mp 135 °C; *H NMR (TFA-d, 360 MHz): dH 8.72 (H-2), 8.19 (H-3), 8.46 (H-4), 7.71 (H-5), 8.06 (H-6), 7.89 (H-7), 9.72 (H-11), 4.02 (H-13), 3.82 (H-14), 3.39 (H-16); 13C NMR (TFA-d, 90 MHz): dC 143.39 (C-2), 110.69 (C-3), 153.13 (C-3a), 116.98 (C-3b), 127.42 (C-4), 126.82 (C-5), 139.34 (C-6), 119.91 (C-7), 141.85 (C-7a), 136.70 (C-8a), 106.18 (C-9), 144.91 (C-9a), 158.05 (C-11), 130.96 (C-12a), 134.13 (C-12b), 121.00 (C-12c), 27.84 (C-13), 56.56 (C-14), 45.26 (15-NCH3); HRFABMS m/z: 333.1189 [M + H] + (Ci9Hi7N4S); Sponges Dercitus sp. and Stetleta sp.18

Dercitamide (72)

fX 12 112b M 3a 4 S 5 / || | 12c II | ^ ^-JL. A 72 ^14 NHCH2CH3 15 23 Yellow solid; mp 192 °C; 1H NMR (TFA-d, 360 MHz): dH 8.66 (H-2), 8.12 (H-3), 8.41 (H-4), 7.66 (H-5), 7.98 (H-6), 7.84 (H-7), 9.88 (H-11), 3.86 (H-13), 3.71 (H-14); 13C NMR (TFA-d, 90 MHz): dC 144.07 (C-2), 111.32 (C-3), 153.70 (C-3a), 117.57 (c-3b), 127.51 (C-4), 128.06 (C-5), 139.95 (C-6), 120.53 (C-7), 142.60 (C-7a), 137.63 (C-8a), 110.43 (C-9), 144.60 (C-9a), 159.88 (C-11), 128.31 (C-12a), 133.46 (C-12b), 121.89 (C-12c), 30.93 (C-13), 39.69 (C-14); HRFABMS m/z: 375.1285 [M + H]+ (C21H19N4S); Sponges Dercitus sp. and Stetleta sp.18

Amphimedine (73) 2 13 Asf J4 N ^N J' H3C 8a^7a"- N 14 73 O 7 Yellow solid; mp >360 °C; UV (EtOH) kmax (e): 210 (19,690), 233 (39,393), 281 (9099), 341 (6060) nm; UV (EtOH + NaBH4) kmax (e): 235 (12,879), 280 (9090) nm; 1H NMR (CDCl3/TFA-d (1:2), 300 MHz): dH 8.68 (d, J = 8.5 Hz, H-1), 7.96 (t, J = 8.5 Hz, H-2), 8.22 (t, J = 8.5 Hz, H-3), 8.97 (d, J = 8.5 Hz, H-4), 9.53 (d, J = 7.0 Hz, H-5), 9.52 (d, J = 7.0 Hz, H-6), 9.20 (s, H-9), 8.52 (d, H-12), 4.10 (s, H-14); 13C NMR (CDCl3/TFA-d (1:2), 75 MHz): dC 133.1 (C-1), 137.4 (C-2), 132.5 (C-3), 125.8 (C-4), 120.5 (C-4a), 146.2 (c-4b), 125.2 (C-5), 139.0 (C-6), 139.8 (c-7a), 175.0 (C-8), 113.5 (C-8a), 147.0 (C-9), 165.9 (c-11), 115.0 (c-12), 143.9 (C-12a), 145.1 (C-12b), 119.0 (C-12c), 147.9 (C-13a), 40.0 (C-14); HRMS m/z: 313.08547 [M]+ (C19HuN3O2); Pacific Ocean sponge Amphimedon sp.54

2-Bromoamphimedine (74) Br 13 Ai \ 4 N H3C ^^ ' N 14 9 74 O 7 A yellow solid; mp >300 °C; UV (MeOH) kmax (e): 236 (23,225), 278 (8680), 309 (8797), 371 (4653) nm; IR (film) vmax: 1680, 1640, 1593 cm"1; 1H NMR (CDCl3, 300 MHz): dH 8.74 (d, J = 1.9 Hz, H-1), 8.04 (dd, J = 8.7, 1.9 Hz, H-3), 8.21 (d, J = 8.7 Hz, H-4), 8.55 (d, J = 5.5 Hz, H-5), 9.30 (d, J = 5.5 Hz, H-6), 8.78 (s, H-9), 8.00 (s, H-12), 3.78 (s, H-14); 1H NMR (CDCl3/TFA-d, 500 MHz): dH 8.84 (d, J = 1.9 Hz, H-1), 8.22 (dd, J = 8.8, 1.9 Hz, H-3), 8.35 (d, J = 8.8 Hz, H-4), 8.95 (brs, H-5), 9.43 (brs, H-6), 8.90 (s, H-9), 8.17 (s, H-12), 3.80 (s, H-14); 13C NMR (CDCl3/TFA-d, 125 MHz): dC 126.9 (C-1), 117.9 (C-2), 138.1 (C-3), 133.7 (C-4), 114.4 (C-4a), 122.0 (C-4b), 122.7 (C-5), 146.0 (c-6), 126.2 (c-7a), 174.7 (C-8), 113.4 (C-8a), 145.7 (C-9), 164.2 (C-11), 115.6 (C-12), 142.8 (C-12a), 142.8 (C-12b), 111.1 (C-12c), 144.9 (C-13a), 39.7 (c-14); ESITOFMS m/z: 392.0035 [M + H]+ (calcd for C19H„O2N3Br 392.0034); Thai marine sponge Petrosia n. sp.44 (continued on next page)

Table 2 (continued)

Demetyldeoxyamphimedine (75)

¿S™ 10

Brown TFA salt; 'H NMR (CDCl3/CD3OD (2:1), 400 MHz): dH 8.20 (d, J = 8.2 Hz, H-1), 7.82 (ddd, J = 8.2, 1.1, 0.8 Hz, H-2), 7.73 (ddd, J = 8.2, 1.1, 0.8 Hz, H-3), 8.53 (d, J = 8.2 Hz, H-4), 8.64 (d, J = 5.3 Hz, H-5), 9.09 (d, J = 5.3 Hz, H-6), 9.39 (s, H-9), 8.86 (d, J = 5.4 Hz, H-11), 8.76 (d, J = 5.4 Hz, H-12); *H NMR (TFA-d/CDCl3 (2:1), 400 MHz): dH 9.16 (d, J =8.1 Hz, H-1), 8.83 (brt, J = 7.69 Hz, H-2), 8.69 (brt, J = 7.69 Hz, H-3), 9.41 (d, J =8.1 Hz, H-4), 9.78 (d, J =6.1 Hz, H-5), 10.08 (d, J =6.1 Hz, H-6), 10.24 (s, H-9), 10.00 (d, J = 6.1 Hz, H-11), 9.86 (d, J = 6.1 Hz, H-12); 13H NMR (CDCl3/CD3OD (2:1), 100 MHz): dC 131.5 (C-1), 132.1 (C-2), 130.3 (C-3), 123.1 (C-4), 121.9 (C-4a), 138.1 (C-4b), 120.6 (C-5), 149.7 (C-6), 145.6 (C-7a), 180.4 (C-8), 126.9 (C-8a), 149.2 (C-9), 151.9 (C-11), 119.4 (C-12), 143.5 (C-12a), 145.7 (C-12b), 118.8 (C-12c), 145.1 (C-13a); 13C NMR (TFA-d/CDCl3 (2:1), 100 MHz): dC 134.7 (C-1), 135.3 (C-2), 138.1 (C-3), 126.1 (C-4), 121.1 (C-4a), 146.7 (C-4b), 126.6 (c-5), 141.4 (C-6), 137.9 (C-7a), 173.5 (c-8), 128.5 (c-8a), 143.9 (C-9), 146.4 (C-11), 124.7 (c-12), 150.3 (C-12a), 143.3 (C-12b), 119.1 (C-12c), 147.8 (C-13a); HRESITOFMS m/z: 306.0642 [M + Na] + (calcd for C18H9N3ONa 330.0638); Western Mediterranean ascidian Cystodytes dellechiajei10

1-Hydroxy-deoxyamphimedine trifluoroacetate salt (76)

Red-orange amorphous solid; UV (MeOH) kmax (log e): 204 (4.51), 246 (4.35), 286 (sh, 4.16), 392 (3.92) nm; IR (film) vmax: 3382, 2925, 2854, 1687, 1598, 1507, 1206, 1136, 801, 723 cm"1; *H NMR (DMSO-d6, 500 MHz): dH 7.48 (d, J = 8.0 Hz, H-2), 7.91 (dd, J = 8.0, 8.0 Hz, H-3), 8.45 (d, J = 8.0 Hz, H-4), 9.13 (d, J = 6.0 Hz, H-5), 9.37 (d, J = 6.0 Hz, H-6), 9.87 (s, H-9), 9.40 (d, J = 6.5 Hz, H-11), 9.90 (d, J = 6.5 Hz, H-12), 4.57 (s, H-14); 13C NMR (DMSO-d6, 125 MHz): dC 156.1 (C-1), 116.5 (C-2), 133.3 (C-3), 114.5 (C-4), 123.8 (C-4a), 137.9 (C-4b), 121.8 (C-5), 150.2 (C-6), 147.0 (C-7a), 180.0 (c-8), 130.3 (C-8a), 146.0 (C-9), 147.8 (C-11), 123.5 (C-12), 147.9 (C-12a), 143.1 (C-12b), 120.1 (C-12c), 134.4 (C-13a), 48.3 (C-14); HRESIMS m/z: 314.0945 [M + H]+ (calcd for C19H12N3O2 314.0930); Palau marine sponge Xestospongia cf. carbonaria45

3-Hydroxy-deoxyamphimedine trifluoroacetate .salt (77)

Neoamphimedine (78)

Orange-yellow amorphous solid; UV (MeOH) kmax (log e): 206 (4.95), 244 (4.94), 394 (3.34), 488 (3.24) nm; IR (film) vmax: 3421, 2925, 1685, 1505, 1443, 1210, 1139, 844, 803, 724 cm"1; 1H NMR (DMSO-d6, 500 MHz): dH 8.39 (d, J = 9.0 Hz, H-1), 7.63 (dd, J = 9.0, 2.5 Hz, H-2), 8.20 (d, J = 2.5 Hz, H-4), 9.00 (d, J = 6.0 Hz, H-5), 9.30 (d, J = 6.0 Hz, H-6), 9.82 (s, H-9), 9.17 (d, J = 6.5 Hz, H-11), 9.38 (d, J = 6.5 Hz, H-12), 4.58 (s, H-14); 13C NMR (DMSO-d6, 125 MHz): dC 135.8 (C-1), 124.3 (C-2), 162.7 (C-3),

108.0 (C-4), 126.6 (C-4a), 140.9 (C-4b), 122.9 (C-5), 151.4 (C-6), 149.5 (C-7a), 180.6 (c-8), 130.8 (C-8a), 147.9 (C-9), 148.7 (C-11), 124.1 (C-12), 149.2 (C-12a), 145.5 (C-12b), 120.8 (C-12c), 141.5 (C-13a), 49.3 (C-14); HRESIMS m/z 314.0938 [M + H]+ (calcd for C19H12N3O2 314.0930); Sponge Xestospongia cf. carbonaria45

Yellow solid; mp >300 °C; FTIR (KBr) vmax: 3100-3000, 1688, 1621, 1606, 1593, 1528, 1340, 1071, 1001, 928, 840, 799, 775, 758, 728 cm"1; UV (EtOH) kmax (e): 205 (26,370), 226 (23,740), 278 (16,870), 371 (10,190) nm; 1H NMR (CDCl3, 500 MHz): dH 8.36 (d, H-1), 7.96 (t, H-2), 7.86 (t, H-3), 8.62 (d, H-4), 8.56 (d, H-5), 9.30 (d, H-6), 7.83 (d, H-11), 7.81 (d, H-12), 3.70 (s, H-14); 1H NMR (CDCl3/CD3OD, 500 MHz): dH 8.14 (H-1), 7.78 (H-2), 7.69 (H-3), 8.48 (H-4), 8.53 (H-5), 8.98 (H-6), 7.87 (H-11), 7.70 (H-12), 3.50 (H-14); 13C NMR (CDCl3/CD3OD, 125 MHz): dC 131.9 (C-1), 131.6 (C-2), 130.2 (C-3), 123.0 (C-4), 121.9 (C-4a), 137.5 (C-4b), 119.0 (C-5), 149.4 (C-6), 146.5 (C-7a), 179.8 (C-8), 118.9 (C-8a), 159.9 (C-9), 145.2 (C-11), 101.7 (C-12), 149.5 (c-12a), 146.2 (C-12b), 117.5 (C-12c), 145.2 (C-13a), 38.0 (C-14); 1H NMR (CDCl3/TFA-d, 360 MHz): dH 8.70 (d, J = 8.0 Hz, H-1), 8.38 (t, J = 8.0 Hz, H-2), 8.24 (t, J = 8.0 Hz, H-3), 8.95 (d, J = 8.0 Hz, H-4), 9.48 (d, J = 6.6 Hz, H-5), 9.33 (d, J = 6.6 Hz, H-6), 8.48 (d, J = 8.0 Hz, H-11), 8.51 (d, J = 8.0 Hz, H-12), 4.06 (s, H-14); 13C NMR (CDCl3/TFA-d, 90 MHz): dC 134.3 (C-1), 137.5 (C-2), 134.3 (C-3), 125.7 (C-4), 120.7 (C-4a), 145.6 (C-4b), 125.5 (C-5), 140.3 (C-6), 138.7 (C-7a), 175.5 (C-8), 116.7 (c-8a), 161.9 (C-9), 144.2 (c-11)

108.1 (C-12), 151.5 (C-12a), 145.0 (C-12b), 117.9 (C-12c), 148.0 (C-13a), 40.2 (C-14); HRFABMS m/z: 314.0948 [M + H]+ (calcd for C19H12N3O2 314.0930); Philippine sponge Xestospongia sp. and Micronesian sponge Xestospongia cf. carbonaria55

Table 2 (continued)

Deoxyamphimedine (79)

Cystodamine (80)

9a 8a" N' 80 O

2-Bromoleptoclinidinone (81)

7a || J4b

Yellow-brown amorphous solid; UV (MeOH) kmax (log e): 206 (3.92), 244 (3.93), 294 (sh, 3.52), 388 (3.51), 478 (2.59) nm; FTIR (film) vmax: 2925, 1684, 1620, 1595, 1510, 1197, 1130 cm"1; 1H NMR (CD3OD, 500 MHz): dH 8.55 (d, J = 6.8 Hz, H-1), 8.13 (dt, J = 7.7, 6.8 Hz, H-2), 8.06 (t, J = 7.7 Hz, H-3), 8.97 (d, J = 7.7 Hz, H-4), 9.13 (d, J = 6.0 Hz, H-5), 9.32 (d, J = 6.0 Hz, H-6), 9.87 (s, H-9), 9.28 (d, J = 6.0 Hz, H-11), 9.45 (d, J = 6.0 Hz, H-12), 4.64 (s, H-14); 13C NMR (CD3OD, 125 MHz): dC 133.3 (C-1), 133.9 (C-2), 132.9 (C-3), 125.2 (C-4), 124.3 (C-4a), 139.9 (C-4b), 122.9 (C-5), 151.4 (c-6),

149.5 (c-7a), 179.2 (C-8), 131.6 (C-8a), 147.9 (C-9), 149.3 (C-11), 124.6 (C-12), 147.3 (C-12a), 145.5 (C-12b), 121.0 (C-12c), 146.7 (C-13a), 49.0 (C-14); 13C NMR (TFA-d/CDCl3 (2:1), 125 MHz): dC 136.4 (C-1), 139.6 (C-2), 136.8 (C-3), 127.2 (C-4), 122.7 (C-4a), 147.8 (c-4b), 127.6 (C-5), 142.8 (C-6), 139.7 (C-7a), 174.9 (C-8), 130.5 (C-8a), 148.8 (C-9), 151.3 (c-11), 128.2 (c-12), 150.5 (C-12a), 144.7 (C-12b), 120.9 (C-12c), 149.4 (C-13a), 51.8 (C-14); HREIMS m/z: 298.0966 [M]+ (calcd for C19H12N3O 298.0980); Sponges Xestospongia sp. and Xestospongia cf. carbonaria56

UV (1N HCl + CH3OH) kmax (log e): 250 (3.62), 278 (3.35), 385 (3.15) nm; IR (CHC13 + TFA-d) mmax: 3025, 1680, 1601, 1462, 1207, 1141, 845 cm"1; 1H NMR (cD2Cl2 + CF3COOD, 400 MHz): dH 8.65 (d, J = 8.0 Hz, H-2), 8.28 (t, J = 8.0 Hz, H-3), 8.26 (t, J = 8.0 Hz, H-4), 8.92 (d, J = 8.0 Hz, H-5), 8.98 (d, J = 5.6 Hz, H-6), 9.35 (d, J = 5.6, H-7), 8.85 (d, J = 6.8, H-11), 7.67 (d, J = 6.8 Hz, H-l 2); 13C NMR (CD2Cl2 + CF3COOD, 100 MHz): dC 145.1 (c-1a), 132.4 (C-2), 134.9 (C-3), 134.7 (C-4),

124.6 (C-5), 125.2 (C-5a), 139.1 (C-5b), 122.4 (C-6), 150.6 (C-7), 140.5 (C-8a), 175.3 (c-9), 148.3 (C-9a), 146.3 (C-11), 118.4 (C-12), 158.0 (C-13); HRFABMS m/z: 273.0884 (C17H„N3O); FABMS m/z (rel. int.): 273 (25), 245 (6.5), 176 (40), 154 (100); Mediterranean ascidian Cystodytes delle chiajei49

Yellow powder; mp >300 °C; UV (EtOH) kmax (e): 371 (21,000), 335 (18,500), 298 (30,600), 278 (32,000), 254 (sh, 27,300), 247 (27,800), 227 (151,600) nm; IR (film) vmax: 3400 (br), 1680, 1600, 1580, 1415, 1270 cm"1; 1H NMR (CDCl3, 300 MHz): dH 8.73 (d, J = 2.1 Hz, H-1), 7.98 (dd, J = 8.7, 2.1 Hz, H-3), 8.49 (d, J = 8.7 Hz, H-4), 9.24 (d, J = 5.7 Hz, H-6), 8.45 (d, J = 5.7 Hz, H-7), 9.15 (dd, J = 4.6, 1.8 Hz, H-9), 7.67 (dd, J = 7.9, 4.6 Hz, H-10), 8.76 (dd, J = 7.9, 1.8 Hz, H-11); 13C NMR (CDCl3, 75 MHz): dC 135.3 (C-1), 126.2 (C-2), 134.0 (C-3), 124.2 (C-4), 122.1 (C-4a), 149.9 (C-4b), 150.1 (C-6), 116.1 (c-7), 137.7 (c-7a), 152.1 (C-7b), 155.6 (C-9), 125.8 (C-10), 136.6 (C-11), 128.9 (C-11a), 181.3 (C-12), 146.7 (C-12a), 117.9 (C-12b), 146.3 (C-13a); LRMS m/z (rel. int.):

363.0 [M + 2H] + (79), 361.0 [M]+ (80), 335.0 [M + 2H-CO]+ (47), 333.0 [M-CO]+ (48),

283.1 (19), 255.1 (33), 254 [M-CO-Br]+ (100); HRMS m/z: 362.98302 [M + H] + (98%) (Cl8H8O81BrN), 360.98507 (100%) C18H8O79BrN3; Ascidian Leptoclinides sp.15

Table 3 Hexacyclic pyridoacridine alkaloids.

Nordehydrocyclodercitin (82)

Stellettamine (83)

Yellow solid; UV/vis (MeOH) kmax (log e): 256 (4.37), 293 (4.11), 338 (3.63), 360 (3.67) nm; IR (CHCl3) mmax: 1603, 1458 cm"1; 1H NMR (CDCl3, 300 MHz): dH 9.28 (d, J = 5.4 Hz, H-2), 8.20 (d, J = 5.4 Hz, H-3), 8.61 (brd, J = 8.3 Hz, H-4), 7.59 (brt, J = 7.6 Hz, H-5), 7.81 (brt, J = 7.6 Hz, H-6), 8.12 (d, J = 8.3 Hz, H-7), 8.29 (d, J = 2.8 Hz, H-9), 7.39 (d, J = 2.8 Hz, H-10), 9.13 (s, H-12); 13C NMR (CDCl3, 100 MHz): dC 148.4 (C-2), 111.1 (C-3), 133.4 (C-3a), 120.6 (C-3b), 125.8 (c-4), 124.8 (C-5), 131.3 (C-6), 115.5 (C-7), 134.5 (C-7a), 117.7 (C-9), 106.8 (C-10), 112.0 (C-10a), 132.8 (C-10b), 149.3 (C-12), 145.3 (C-13a), 141.8 (C-13b), 113.1 (C-13c), 124.8 (C-13d); HRESIMS m/z: 300.0587 [M + H] + (Calcd for C18H10N3S+ 300.0589); Australian marine ascidian Aplidium sp., cf. Aplidium cratiferum57

Dark brown roughly cubic crystals; mp 280-282 °C; UV (MeOH) kmax (log e): 205 (4.197), 260 (3.973), 309 (4.032) 338 (3.570), 364 (3.575), 391 (3.534), 460 (3.136) nm; IR (CHCl3) vmax: 1690, 1658, 1629, 1590, 1535, 1450, 1428, 1345, 1260,1220 cm"1; HRFABMS m/z: 365.0839 (calcd for C20H14N4S + Na 365.0837), 343.1039 (calcd for C20H15N4S 343.1019);10 1H NMR (CDCl3, 300 MHz): dH 9.16 (d, J = 5.3 Hz, H-2), 8.10 (d, J = 5.3 Hz, H-3), 8.55 (dd, J = 8.3, 1.9 Hz, H-4), 7.51 (ddd, J = 8.0, 7.9, 1.4 Hz, H-5), 7.67 (ddd, J = 8.6, 7.9, 1.8 Hz, H-6), 9.18 (dd, J = 8.6, 1.4 Hz, H-7), 6.86 (s, H-10), 9.06 (s, H-12), 2.95 (s, NCH3); 13C NMR (CDCl3, 100 MHz): dC 147.5 (C-2), 110.5 (C-3), 133.0 (C-3a), 121.4 (C-3b), 124.3 (C-4), 124.8 (C-5), 130.8 (C-6), 118.4 (C-7), 136.4 (C-7a), 149.4 (C-9), 94.2 (C-10), 110.6 (C-10a), 129.7 (C-10b), 149.1 (C-12), 145.0 (C-13a), 140.8 (C-13b), 113.3 (C-13c), 122.0 (C-13d), 45.3 (NCH3); Australian marine ascidian Aplidium sp., cf.

Aplidium cratiferum19,57

(continued on next page)

Table 3 (continued)

N—CH3

Cycloshermilamine D (84)

^¿k Yellow amorphous powder; UV (MeOH) kmax (log e): 208 (4.18), 215 (4.16), 262 (4.04), 299 (3.88), 391 (3.35),

H L T 4 480 (3.19) nm; IR (neat) mmax: 1682, 1601, 1560, 1462, 1336, 1221 cm"1; UV (MeOH, H+) kmax (log e): 214

o^13 „^(4.49), 288 (4.06), 427 (3.4) nm; 1H NMR (CDCl3, 500 MHz): dH 8.83 (d, J = 5.2 Hz, H-2), 7.96 (d,

J = 5.2 Hz, H-3), 8.47 (brd, J = 8.0 Hz, H-4), 7.55 (brt, J = 7.5 Hz, H-5), 7.80 (brt, J = 7.5 Hz, H-6), 9.13 (brd, J = 8.0 Hz, H-7), 6.78 (brs, H-10), 3.64 (brs, H-12), 9.25 (brs, H-14), 2.96 (brs, NMe); 13C NMR (cDCl3, 125 MHz): dC 143.6 (C-2), 109.2 (C-3), 131.5 (C-3a), 120.5 (C-3b), 125.7 (C-4), 124.6 (C-5), 132.5 (C-6), 118.5 (C-7), 136.5 (C-7a), 150.1 (C-9), 94.3 (C-10), 115.0 (C-10a), 113.8 (C-10b), 29.6 (C-12), 163.4 (C-13), 120.3 (C-14a), 137.3 (C-14b), 112.6 (C-14c), 118.8 (C-14d), 45.2 (NMe); EIMS m/z: 372 [M] + (100), 357 (20), 330 (20), 288 (14); HREIMS m/z: 372.1045; Tunicate Cystodytes violatinctus58

13-Didemethylaminocycloshermilamine D (85)

Yellow-brown TFA salt; 1H NMR (DMSO-d6, 400 MHz): dH 8.55 (d, J = 8.2 Hz, H-1), 7.93 (brd, J = 7.9 Hz, H-2), 7.67 (brd, J = 7.9 Hz, H-3), 8.83 (d, J = 8.2 Hz, H-4), 8.43 (d, J = 5.3 Hz, H-5), 9.08 (d, J = 5.3 Hz, H-6), 3.77 (brs, H-10), 7.30 (d, J = 2.4 Hz, H-12), 8.86 (d, J = 2.4 Hz, H-12a); 1H NMR (CDCl3, 400 MHz): dH 8.12 (d, J = 8.2 Hz, H-1), 7.86 (brd, J = 7.9 Hz, H-2), 7.62 (brd, J = 7.9 Hz, H-3), 8.57 (d, J = 8.2 Hz, H-4), 8.09 (d, J = 5.3 Hz, H-5), 9.03 (d, J = 5.3 Hz, H-6), 9.56 (brs, H-8), 3.77 (brs, H-10), 7.29 (brd, H-12), 8.27 (brd, H-12a), 8.27 (d, J = 2.4 Hz, H-13); 13C NMR (DMSO-d6, 100 MHz): dC 117.5 (C-1), 125.9 (C-2), 133.1 (C-3), 127.7 (C-4), 120.7 (C-4a), 132.9 (C-4b), 115.5 (C-4d), 112.5 (C-5), 148.7 (C-6), 137.3 (C-7a), 164.5 (C-9), 29.9 (C-10), 114.5 (C-11a), 113.8 (C-11b), 112.3 (C-12), 121.3 (C-13), 135.5 (C-14a); 13C NMR (CDCl3, 100 MHz): dC 114.8 (C-1), 131.3 (C-2), 124.0 (C-3), 125.0 (C-4), 119.2 (C-4a), 133.5 (C-4b), 121.3 (C-4d), 109.5 (C-5), 146.0 (C-6), 137.3 (C-7a), 164.5 (C-9), 29.9 (C-10), 114.5 (C-11a), 119.2 (C-11b), 106.5 (C-12), 117.5 (C-13), 134.0 (C-14a); HRESITOFMS m/z: 330.0697 [M + H] + (calcd for Ci9Hi2N3OS, 330.0696); Western Mediterranean ascidian Cystodytes dellechiajei10

Sebastianine B (86)

Segoline A (87)

HjCa « 11

"I H I» -M

h^^^ * ch3

I CH3I

O^ N^O

(log e): 206 (4.57), 252 (4.58), 292 (4.23), 366 (4.19) nm; IR

Pale yellow amorphous solid; UV (MeOH) k, (KBr) mmax: 3400 (br), 1650, 1595, 1411, 1366, 1037, 756 cm"1; 1H NMR (DMSO-d6, 500 MHz): dH 8.08 (d, J = 7.8 Hz, H-1), 7.84 (t, J = 7.8 Hz, H-2), 7.69 (t, J = 7.8 Hz, H-3), 8.76 (d, J = 7.8 Hz, H-4), 8.74 (d, J = 5.6 Hz, H-5), 9.03 (d, J = 5.6 Hz, H-6), 4.04 (ddd, J = 12.0, 12.0, 7.5 Hz, H-10A), 4.20 (ddd, J = 12.0, 12.0, 5.7 Hz, H-10B), 3.14 (ddd, J = 12.0, 10.1, 7.5 Hz, H-11A), 3.24 (ddd, J = 12.0, 10.1, 5.7 Hz, H-11B), 4.72 (s, H-14), 2.50 (m, H-15), 1.12 (d, J = 7.0 Hz, H-16), 0.82 (d, J = 7.0 Hz, H-17), 7.92 (s, OH); 13C NMR (DMSO-d6, 125 MHz): dC 129.2 (C-1), 131.4 (C-2), 127.0 (C-3), 123.9 (C-4), 120.8 (C-4a), 137.0 (C-4b), 116.8 (c-5), 148.5 (C-6), 156.0 (C-7a), 86.6 (C-8), 145.4 (C-8a), 45.4 (C-10), 25.7 (C-11), 114.2 (C-11a), 149.7 (C-11b), 114.5 (C-11c), 145.1 (C-12a), 165.7 (C-13), 76.6 (C-14), 29.1 (C-15), 19.2 (C-16), 16.4 (C-17); EIMS m/z: 373.1428 [M]+ (calcd for C22H19N3O3 373.1426); Brazilian ascidian Cystodytes dellechiajei48

Mp >276 °C; [a]D: -322 (c 1.0, CHCl3); UV (MeOH) kmax (e): 460 (3100), 383 (2600), 368 (1600), 320 (5600), 308 (5100), 274 (16,200), 236 (9000s) nm; UV (MeOH, H+)) kmax (e): 545 (2500), 382 (2500), 366 (1400), 298 (21,300), 278 (15600s), 245 (7400) mn; IR (CHCl3) vmax: 2840, 1710, 1585, 1520, 1400, 1290, 1140 cm-1; Tunicate Eudistoma sp.;39 [a]D -325; 1H NMR (CDCl3, 400 MHz): dH 8.75 (d, J = 5.0 Hz, H-2), 7.48 (d, J = 4.8 Hz, H-3), 7.93 (d, J = 8.5 Hz, H-4), 7.11 (dt, J = 7.9 Hz, H-5), 7.34 (dt, J = 7.8 Hz, H-6), 7.70 (d, J = 8.6 Hz, H-7), 3.75 (d, J = 2.0 Hz, H-13), 6.95 (s, H-14), 2.00 (dq, J = 7.3, 2.2 Hz, H-16), 1.88 (s, H-17), 1.31 (d, J = 7.0 Hz, H-18), 4.05 (s, H-19); 13C NMR (CDCl3, 100 MHz): dC 149.6 (C-2), 108.6 (C-3), 138.8 (C-3a), 120.9 (C-3b), 124.4 (C-4), 121.9 (C-5), 130.1 (C-6), 118.6 (C-7), 139.2 (C-7a), 61.6 (C-9), 172.9 (C-10), 170.6 (C-12), 49.9 (C-13), 111.2 (C-13a), 127.3 (C-13b), 109.2 (C-14), 147.2 (C-15), 139.9 (C-15a), 120.5 (C-15b), 37.1 (C-16), 18.5 (C-17), 15.1 (C-18), 55.6 (C-19); Indian Ocean tunicate Eudistoma bituminis6

Isosegoline A (88)

H3CO. 1'

■ JÎ CH3 HN-7v 3 12 11 xO

[a]D -660 (c 0.001, CHCl3); UV (MeOH) kmax (e): 470 (3500), 386 (3100), 360 (2300), 330 (9400), 274 (19,100), 242 (13,900) nm; UV (MeOH, H+) kmax (e): 550 (7800), 388 (7700), 368 (6100), 297 (25,000), 248 (13,400) nm; IR (CHC13) vmax: 2850, 1730, 1580, 1520, 1280, 1150 cm-1; 1H NMR (DMSO-d6, 360 MHz): dH 8.57 (d, J = 4.8 Hz, H-2), 7.63 (d, J = 4.8 Hz, H-3), 7.47 (d, J = 7.9 Hz, H-4), 7.06 (t, J = 7.9 Hz, H-5), 7.48 (t, J = 7.9 Hz, H-6), 8.08 (d, J = 7.9 Hz, H-7), 5.06 (q, J = 6.5 Hz, H-9), 11.20 (brs, 12-NH), 3.88 (s, H-14), 7.18 (s, H-15), 1.04 (d, J = 6.5 Hz, 17-CH3), 1.45 (s, 18-CH3), 3.89 (s, 19-CH3); 13CNMR (CDCl3, 90 MHz): dC 150.9 (C-2), 109.9 (C-3), 147.8 (C-3a), 118.9 (C-3b), 124.4 (c-4), 121.4 (C-5), 132.4 (C-6), 114.5 (C-7), 140.5 (C-7a), 51.1 (C-9), 52.7 (C-10), 180.5 (C-11), 175.2 (C-13), 49.3 (C-14), 107.4 (C-14a), 138.9 (C-14b), 108.8 (C-15), 143.1 (C-16), 127.0 (C-16a), 120.4 (C-16b), 12.3 (C-17), 23.6 (C-18), 56.5 (C-19); CIMS (methane) mjz (rel. int.): 386 [M + H]+ (C23H20N3O3, 100); Tunicate Eudistoma sp.39,40

Table 3 (continued)

Segoline B (89)

13 CH3 9_„.. h»-4.„(j ch3

(S>| \(S) 17

O*^ N^* O

Segoline C (90)

19 H3CO.

H^jxgsT CH3

(S)ltiSj iS> 17

Cyclodercitin (91) H3C^ * 2

[a]D +355 (c 1, CHCl3); UV (MeOH) kmax (e): 460 (4200), 383 (3700), 368 (2500), 320 (7400), 310 (7700), 265

(e): 544 (4800), 383 (3600), 368 (2100), 298 (31,300), 280

(24,900), 242 (11,300) nm; UV (MeOH, H +

(24,000), 245 (11,300) nm; IR (CHCl3) vmax: 3700, 3620, 3040, 1710, 1520, 1220, 1050, 940, 880 cm"1; 1H NMR (DMSO-d6, 360 MHz): dH 8.64 (H-2), 7.74 (H-3), 7.80 (d, J = 4.8 Hz, H-4), 7.43 (H-5), 7.13 (H-6), 8.12 (H-7), 11.47 (brs, 11-NH), 3.80 (H-13), 7.14 (H-14), 2.83 (H-16), 1.77 (H-17), 0.68 (H-18), 3.92 (H-19); 13C NMR (CDCl3, 90 MHz): dC 144.0 (C-2), 107.6 (C-3), 146.9 (C-3a), 118.5 (C-3b), 125.8 (C-4), 123.6 (C-5), 133.5 (C-6), 119.8 (C-7), 140.1 (C-7a), 130.5 (C-8a), 61.6 (C-9), 170.4 (C-10), 171.6 (C-12), 47.9 (C-13), 110.5 (C-13a), 115.4 (C-14), 141.8 (C-15), 128.8 (C-15a), 120.0 (C-15b), 34.5 (C-16), 18.2 (C-17), 12.4 (C-18), 56.4 (c-19); CIMS (CH4) m/z (re1. int.): 386 [M + H]+ (C23H20N3O3, 100); Tunicate Eudistoma sp.40

[a]D -374; 'H NMR (CDCl3, 400 MHz): dH 8.76 (d, J = 4.9 Hz, H-2), 7.50 (d, J = 5.0 Hz, H-3), 7.91 (dd, J = 8.0, 1.4 Hz, H-4), 7.10 (brt, J = 7.8 Hz, H-5), 7.35 (ddd, J = 8.8, 7.2, 1.6 Hz, H-6), 7.82 (brd, J = 8.7 Hz, H-7), 11.5 (11-NH), 3.69 (d, J = 3.0 Hz, H-13), 6.97 (s, H-14), 2.54 (dq, J = 6.7, 3.1 Hz, H-16), 1.92 (s, H-17), 0.87 (d, J = 6.7 Hz, H-18), 4.06 (s, H-19); 13C NMR (CDCl3, 100 MHz): dC 150.8 (C-2), 109.2 (C-3), 138.4 (c-3a), 121.2 (C-3b), 124.5 (c-4), 122.5 (C-5), 130.6 (C-6), 119.2 (C-7), 140.0 (C-7a), 60.4 (C-9), 172.3 (C-10), 170.4 (C-12), 48.5 (C-13), 106.4 (C-13a), 128.3 (C-13b), 110.2 (C-14), 148.3 (c-15), 141.1 (C-15a), 120.6 (C-15b), 35.7 (C-16), 18.6 (C-17), 13.4 (C-18), 56.3 (C-19); ESIMS m/z: 386 [M + H] + (C23H19N3O3); Tunicate Eudistoma bituminis6

Blue powder; mp 298 °C; 1H NMR (TFA-d, 360 MHz): dH 8.39 (H-2), 7.67 (H-3), 8.26 (H-4), 7.35 (H-5), 7.82 (H-6), 7.32 (H-7), 9.18 (H-11), 3.76 (H-13), 4.40 (H-14), 4.32 (11-NCH3); 13C NMR (TFA-d, 90 MHz): dC 150.49 (C-2), 108.49 (C-3), 140.97 (C-3a), 117.44 (C-3b), 127.71 (C-4), 124.24 (C-5), 137.45 (C-6), 116.03 (C-7), 149.97 (C-7a), 141.06 (C-8a), 116.25 (C-9), 140.81 (C-9a), 151.47 (C-11), 136.65 (C-12a), 134.05 (C-12b), 118.88 (C-12c), 29.09 (C-13), 49.02 (C-14), 51.87 (11-NCH3); HRFABMS m/z: 316.0916 [M + H] + (Ci9Hi4N3S); Marine sponge Dercitus sp.18

such as methyl amino, dimethyl amino, acyl amino, N-ethyl-b, b-dimethylacryloyl amide (as in 3), N-ethyl-a,b-dimethylacryloyl amide (tigloyl, as in 4), fatty acyl ester, and (dimethyl amino)thioethyl.3 Moreover, they showed different ring substituents as thiomethyl, amino, bromide, car-boxymethyl, hydroxyl, and ether functionalities.

Pyridoacridines vary in structure by appendage of different side chains or fusion of rings to ring C (1), and occasionally to the acridine nitrogen to produce different classes: tetracyclic, pentacyclic, hexacyclic, heptacyclic, and octacyclic. The substitution of pyridoacridines with halogen is rare. If it is present, it is mainly with bromine atom at C2 in ring A. The rings oxidation states are variable. In some compounds, ring D is partially saturated. Amphimedine was the first identified pyridoacridine in 1983.3 4 Pyridoacridines from marine sources are classified into five main groups namely, tetracyclic (four rings), penta-cyclic (five rings), hexacyclic (six rings), heptacyclic (seven rings), and octacyclic (eight rings).

Pyridoacridines possess diverse biological activities such as anti-viral,5-8 antimicrobial,9-13 immunosuppressant,14 antiparasitic,4'15 anticholinesterase,16 cytotoxic,14,15,17-27 and insecticidal.28 In addition, they produce reactive oxygen species, inhibit topoisomerases and aspartate semialdehyde dehy-drogenase, cause the release of calcium from the sarcoplasmic reticulum, bind nucleotide receptors, and induce neuronal dif-ferentiation.28,29 The cytotoxic activity of pyridoacridines is due to a highly electron-deficient planar aromatic system that

can intercalate DNA leading to inhibition of cell growth.1,30 During last decades, pyridoacridines and their analogs have represented excellent goals for synthetic researches,30 proving that these alkaloids are an interesting source for the development of new drug leads. Some review articles on biological activities and the structural relatedness of members of pyri-doacridines are available.1,3,4,30 There are currently no reviews pertaining to the structural elucidation of naturally occurring marine pyridoacridines. A review of the available NMR literature for this important class of natural products was therefore considered to be timely. Also, it would appear to be valuable to provide an easy access to an extensive list of spectroscopic and physical data (physical state, melting point (C0), and optical rotation (concentration, solvent)) of pyridoacridines. This review is mainly a compilation of our earlier review, which discussed this class of naturally occurring alkaloids in term of their occurrence, biosynthesis, biological activities, and structural assignment.5 This work provided a summary of the isolation, physical properties, spectral data, sources, and associated references of 95 pyridoacridines. These data have been listed for each compound: name, structure formula, melting point, optical rotation, UV (kmax nm, solvent, log e), IR (absorption bands in cm-1), 1H and 13C NMR (solvent, spectrometer frequency, d values ppm), source, molecular weight, molecular formula, and reference. The 1H and 13C NMR have been written to the second and first decimal points, respectively (Tables 1-4). Pyridoacridines described in this review have

Table 4 Heptacyclic and octacyclic pyridoacridine alkaloids.

Eilatin (92)

Bright yellow; mp >310 °C; UV (MeOH) kmax (e): 242 (48,200), 286 (36,700), 366 (11,500), 388 (21,000), 408 (30,400), 434 (27,000) nm; UV (MeOH, H+) Xmax (e): 305 (39,800), 440 (21,500) nm, UV (MeOH + Ni+2) kmax (e): 296 (38,700), 368 (7300), 404 (18,300), 426 (22,500), 450 (19,900) nm; IR (CHC13) mmax: 3000, 1240, 1200, 1120, 970 cm-1; 'H NMR (CDCl3): dH 8.68 (d, J = 7.2 Hz, H-1), 7.87 (t, J = 7.2 Hz, H-2), 8.00 (t, J = 8.0 Hz, H-3), 8.70 (d, J = 8.0 Hz, H-4), 8.57 (d, J = 5.5 Hz, H-11), 9.32 (d, J = 5.5 Hz, H-12); 13C NMR (CDCl3): dC 132.1 (C-1), 129.3 (C-2), 131.7 (C-3), 122.5 (C-4), 146.1 (C-4a), 150.2 (C-5a), 118.7 (C-5c), 122.3 (C-10a), 148.8 (C-10b), 117.1 (C-11), 149.7 (C-12), 138.8 (C-13a); HREIMS m/z: 356.1062 (C24H12N4); Tunicate Eudistoma sp.7,40

Octacyclic pyridoacridine alkaloids Biemnadin (93)

Eudistone A (94)

Eudistone B (95)

°C; UV (MeOH) kmax (e): 229 (29,000), 256 (17,000), 370 (5000) nm; IR (KBr) mmax: *H NMR (DMSO-d6/CD3OD, 2:1): dH 7.87 (d, J =8.1 Hz, H-l), 7.75 (dd, J =8.1,

J = 5.5 Hz,

Yellow crystal; mp >300 ' 3400, 3250, 1660, 1560 cm"

7.4 Hz, H-2), 7.82 (dd, J = 7.7, 7.4 Hz, H-3), 8.78 (d, J = 7.7 Hz, H-4), 8.80 (d, J = 5.5 Hz, H-5), 9.19 (d, J = 5.5 Hz, H-6), 4.03 (ddd, J = 15.5, 5.1, 1.4 Hz, H-9A), 3.83 (ddd, J = 15.5, 5.1, 1.4 Hz, H-9B), 3.12 (m, H-10A), 2.91 (m, H-10B), 3.79 (m, H2-12A), 3.35 (m, H-12B), 7.87 (dd, J = 8.1, 1.5 Hz, H-14), 6.68 (ddd, J = 8.1, 7.3, 1.1 Hz, H-15), 7.11 (ddd, J = 8.4,7.3, 1.5 Hz, H-16), 6.37 (dd, J = 8.4, 1.1Hz, H-17); l3C NMR (DMSO-d6/ CD3OD, 2:1): dC 128.6 (C-1), 132.2 (C-2), 129.4 (C-3), 124.1 (c-4), 121.5 (C-4a), 137.7 (C-4b), 114.7 (C-4c), 120.2 (C-5), 148.8 (C-6), 144.5 (C-7a), 155.1 (C-7b), 38.3 (C-9), 29.1 (C-10), 162.9 (C-10a), 117.8 (C-10b), 40.6 (C-12), 44.9 (C-12a), 191.3 (C-13), 117.8 (C-13a), 125.7 (C-14), 117.8 (C-15), 135.0 (C-16), 115.9 (C-17), 145.5 (C-17a), 54.7 (C-18a), 154.1 (C-18b), 143.5 (C-19a); HRFABMS m/z: 430.1657 [M + H]+ (calcd for C27H20N5O 430.1668); Okinawan marine sponge Biemna sp.51

Yellow amorphous solid; 1H NMR (TFA salt in DMSO-d6, 500 MHz,): dH 7.69 (d, J = 7.9 Hz, H-1), 7.85 (dd J = 7.9, 7.3 Hz, H-2), 7.76 (dd, J = 7.9, 7.3 Hz, H-3), 8.81 (d, J = 7.9 Hz, H-4), 8.95 (d, J = 5.5 Hz, H-5), 9.17 (d, J = 5.5 Hz, H-6), 10.30 (br, H-8), 3.95 (m, H-9A), 3.64 (m, H-9B), 2.99 (m, H-10A), 2.91 (m, H-10b), 3.71 (1H, m, H-12), 3.58 (dd, J = 12.8, 13.4 Hz, H-12), 3.27 (dd, J = 12.8, 4.9 Hz, H-12a), 7.74 (d, J = 7.3 Hz, H-14), 6.62 (t, J = 7.3 Hz, H-15), 7.10 (t, J = 7.3 Hz, H-16), 6.35 (d, J = 7.3 Hz, H-17), 7.41 (s, H-18); 13C NMR (TFA-salt in DMSO-d6, 125 MHz): dH 129.3 (C-1), 132.1 (C-2), 128.5 (C-3), 124.1 (C-4), 121.2 (C-4a), 137.5 (C-4b), 114.5 (C-4c), 120.2 (C-5), 148.7 (C-6), 144.3 (C-7a), 153.9 (C-7b), 38.2 (c-9), 26.5 (C-10), 162.7 (C-10a), 95.3 (C-10b), 40.5 (C-12), 44.6 (C-12a), 190.9 (C-13), 125.6 (C-14), 117.6 (C-13a, 15), 134.9 (C-16), 115.8 (C-17), 145.3 (C-17a), 54.4 (C-18a), 155.1 (C-18b), 143.2 (C-19a); FABMS m/z: 430 [M + H] + ; Indonesian marine sponge Biemna fortis26

Yellow amorphous powder; UV (MeOH) kmax (e): 210 (44,750), 238 (48,475), 260 (sh, 24,670), 323 (15,320), 338 (15,480), 359 (sh, 13,060), 395 (10,640) nm; IR (neat, AgCl plate) vmax: 3360, 3220, 1660, 1595, 1535, 1200 cm-1; 1H NMR (DMSO-d6/CDCl3, 2:1): dH 8.20 (brd, J = 8.0 Hz, H-1), 7.93 (brt, J = 8.0 Hz, H-2), 7.79 (brt, J = 7.5 Hz, H-3), 8.72 (brd, J = 8.0 Hz, H-4), 8.52 (d, J = 5.5 Hz, H-5), 8.48 (d, J = 5.5 Hz, H-6), 6.34 (d, J = 8.0 Hz, H-9), 7.04 (brt, J = 7.0 Hz, H-10), 6.55 (brt, J = 5.5 Hz, H-11), 7.64 (brd, J = 7.5 Hz, H-12), 2.92 (dd, J = 12.0, 5.5 Hz, H-13a), 3.61 (dd, J = 16.5, 12.0 Hz, H-14ax), 3.74 (m, H-14eq), 2.69 (m, H-16ax), 2.69 (m, H-16eq), 3.59 (dt, J = 16.0, 6.5 Hz, H-17ax), 3.75 (dd, J = 16.0, 5.5 Hz, H-17eq); l3C NMR (DMSO-d6/ CDCl3, 2:1); dC 131.0 (C-1), 132.4 (C-2), 130.5 (C-3), 124.3 (C-4), 123.6 (C-4a), 137.8 (C-4b), 113.7 (C-4c), 116.7 (C-5), 148.0 (C-6), 157.9 (C-7a), 55.0 (C-7b), 146.4 (C-8a), 116.0 (C-9), 135.4 (C-10), 117.9 (C-11), 126.3 (C-12), 191.8 (C-13), 45.7 (C-13a), 41.4 (C-14), 163.8 (C-15a), 114.9 (C-15b), 27.4 (C-16), 38.2 (C-17), 149.9 (C-18a), 144.6 (C-18b), 144.4 (C-19a); HRFABMS m/z: 430.1677 [M + H]+ (calcd for C27H19N5O 430.1668); Seychelles tunicate Eudistoma sp.8

1H NMR (DMSO-d6/CD3OD, 2:1): dH 8.33 (brd, J = 8.0 Hz, H-l), 8.01 (brt, J = 7.5 Hz, H-2), 7.88 (brt, J = 7.5 Hz, H-3), 8.79 (brd, J = 7.5 Hz, H-4), 8.57 (s, H-5), 8.57 (s, H-6), 6.32 (d, J = 8.5 Hz, H-9), 7.11 (brt, J = 8.0 Hz, H-10), 6.62 (brt, J = 8.0 Hz, H-11), 7.68 (dd, J = 8.0, 1.0 Hz, H-12); Okinawan marine sponge Biemna sp.51

White amorphous powder; [a]D -177.8 (c 0.036, MeOH); UV (MeOH) kmax (e): 204 (45,650), 239 (47,365), 259 (39,550), 324 (14,670) nm; IR (neat, AgCl plate) vmax: 3390, 1595, 1515, 1020 cm-1; 1H NMR (DMSO-d6/ CD3OD, 2:1): dH 8.37 (brd, J = 8.5 Hz, H-1), 7.97 (brt, J = 8.0 Hz, H-2), 7.82 (brt, J = 7.5 Hz, H-3), 8.77 (brd, J = 8.0 Hz, H-4), 8.59 (s, H-5), 8.59 (s, H-6), 6.39 (d, J = 8.0 Hz, H-9), 7.13 (brt, J = 7.5 Hz, H-10), 6.65 (brt, J = 7.5 Hz, H-11), 7.71 (dd, J = 8.0, 1.0 Hz, H-12), 3.32 (dd, J = 12.0, 5.5 Hz, H-13a), 3.43 (t, J = 12.0 Hz, H-14ax), 3.55 (dd, J = 12.0, 5.5 Hz, H-14eq), 6.84 (d, J = 6.0 Hz,, H-16), 8.30 (d, J = 6.0 Hz, H-17); 1H NMR (DMSO-d6/CDCl3, 2:1): dH 8.35 (brd, J = 8.0 Hz, H-1), 7.89 (brt, J = 7.5 Hz, H-2), 7.74 (brt, J = 7.5 Hz, H-3), 8.68 (brd, J = 8.0 Hz, H-4), 8.53 (d, J = 6.0 Hz, H-5), 8.50 (d, J = 6.0 Hz, H-6), 6.35 (d, J = 8.0 Hz, H-9), 7.03 (brt, J = 7.0 Hz, H-10), 6.56 (brt, J = 5.5 Hz, H-11), 7.66 (brd, J = 7.5 Hz, H-12), 3.19 (dd, J = 12.0, 6.0 Hz, H-13a), 3.38 (t, J = 12.0 Hz, H-14ax), 3.42 (dd, J = 12.0, 6.0 Hz, H-14eq), 6.69 (d, J = 6.0 Hz,, H-16), 8.32 (d, J = 6.0 Hz, H-17), 7.21 (brs, 8-NH), 7.48 (br, 15-NH); l3C NMR (DMSO-d6/CD3OD, 2:1): dC 130.7 (C-1), 132.2 (C-2), 129.5 (C-3), 124.4 (C-4), 123.0 (C-4a), 137.9 (C-4b), 113.6 (C-4c), 116.7 (C-5), 147.8 (C-6), 157.7 (C-7a), 55.6 (C-7b), 145.26 (C-8a), 116.2 (C-9), 135.3 (C-10), 118.17 (C-11), 126.2 (C-12), 118.20 (C-12a), 191.8 (C-13), 43.9 (C-13a), 40.1 (C-14), 153.4 (C-15a), 114.9 (C-15b), 109.6 (C-16), 146.9 (C-17), 149.3 (C-18a), 145.28 (C-18b), 145.18 (C-19a); HRFABMS m/z: 428.1536 (C27H17N5O 428.1511); Seychelles tunicate Eudistoma sp.8

been arranged in five different groups according to the number of fused rings in their structure, including group I-tetracyclic, group II pentacyclic, group III-hexacyclic, group IV-heptacyclic, and group V-octacyclic (see Figs. 2-5).

2. Isolation and structural characterization of pyridoacridines

The extraction was carried out using MeOH, MeOH/CH2Cl2 (1:1) or MeOH/CH2Cl2/TFA (50:50:0.1). The extract was partitioned between n-hexane, CH2Cl2, and EtOAc. The CH2Cl2 and EtOAc fractions were subjected to successive chromatographies on silica gel, Sephadex LH-20, reversed phase RP-18, Diaion HP-20 or ODS flash chromatography using different solvent mixtures with increasing polarity. The isolated

Pantherinine (1)

Cystodytin A (3)

Cystodytin C (5)

Pantherinine acetate (2)

Cystodytin B (4)

Cystodytin D (6)

Cystodytin I (11) N*

Cystodytin K (13)

CF3CO2 +

NH3O2CCF3

Styelsamine A (15)

CF3CO2 Hv +

Styelsamine C (17)

Diplamine (19)

Cystodytin J (12)

NH3 O2CCF3

Iminoquinone (14) cf3cö2h,*

Styelsamine B ( 16)

CF3CO2 H^f

NH3O2CCF3

Styelsamine D ( 18)

Isodiplamine (20)

Cystodytin E (7)

Cystodytin F (8)

Fig. 2 (continued)

Cystodytin G (9)

Cystodytin H (10)

Figure 2 Tetracyclic pyridoacridine alkaloids 1-26.

compounds can be further purified by preparative HPLC. HPLC has been proved as the best technique for separation, identification, and quantification of pyridoacridines.

2.1. Ultraviolet visible spectroscopy (UV)

Pyridoacridines are colored natural products, having a highly conjugated system. So, ultraviolet visible spectroscopy tech-

Diplamine B (21)

(H3C)2N

(H3C)2N

Tintamine (23)

Varamine A (25)

COOCH3

Nor-segoline (22)

OH (H3C2N

Tintamine diacetate (24)

Varamine B (26)

Fig. 2 (continued)

nique is useful for their identification. The UV spectra in MeOH or EtOH showed absorption bands at kmax 210-390 nm, indicating a polyheteroaromatic system. The a, b-unsaturated ketone moiety was confirmed by the significant changes, that were observed upon addition of NaBH4. A bathochromic shift was observed in the UV spectrum upon addition of acid as trifluoroacetic acid (TFA) or alkaline as KOH or NaOH.

2.2. Infrared spectroscopy (IR)

The presence of a conjugated ketone carbonyl and an amide group is easily detectable in IR spectra as a band in the region of 1640 and 1660 cm-1. The bands at 3370-3215 cm-1 were attributed for primary or secondary amines. Absorption bands due to hydroxyl groups appear at 3400-3600 cm-1.

2.3. NMR spectroscopy

NMR spectral data were obtained in CDCl3, DMSO-d6 or CD3OD. In certain cases, trifluoroacetic acid-d4 was added to the previously mentioned NMR solvents in a ratio 2:1 to

overcome the poor solubility. In the 1H NMR spectra, the di-substituted benzene (ring A) gives rise to four coupled protons (H-1 to H-4, 7.0-9.0 ppm, J = 8-9 Hz). H-1 resonated at downfield due to the acridine nitrogen's deshielding effect. An AB system for tri-substituted pyridine ring at dH 8.5 (d, J = 5.6 Hz, H-5) and 9.0 (d, J = 5.6 Hz, H-6) was observed. 1H and 13C NMR spectra of the majority of alkyl chain substituted pyridoacridines possessed resonances for a free or substituted 2-ethyl amino (NHCH2CH2) side chain at dH 3.08-3.29/ dC 28.2-31.2 and dH 3.30-3.52/dC 36.2-40.2. Moreover, p,p-dimethylacryloyl amide moiety was proved by the observed signals at dH 5.50-5.65 (brs or dd, J = 1.3-1.5 Hz, C—H)/dC 117.4-118.5 (C—H), 1.62-2.00/19.4-20.0 (CH3), 1.93-2.16/26. 4-27.3 (CH3), 150.3-153.5 (qC), and 167.8-169.5 (C=O). However, the a,b-dimethylacryloyl amide moiety was evident by the resonances at dH 6.19-6.48 (m or dd, J = 6.7-7.2, 1.3-1.5 Hz, C-H)/dC 130.0-133.7, 1.60-1.82/11.9-14.2 (CH3), 1.51-1.75/12.2-13.9 (CH3), 130.1-131.3 (qC), and 169.0-171.5 (C=O). The resonances at 179.4-183.8 and 167.8-170.3 ppm indicated the presence of quinoid and amide carbonyls, respectively. The thiomethyl group was evident by the singlet downfield methyl proton at 2.50-2.97 ppm and its corresponding carbon at 16.4-18.7 ppm. The characteristic

Kuanoniamine A (27)

Dehydrokuanoniamine B (29)

Kuanoniamine D (31)

Kuanoniamine F (33)

Kuanoniamine B (28)

Kuanoniamine C (30)

.V-Deacetylkuanoniamine C (32)

Dehydrokuanoniamine F (34)

Sagitol (35) Sagitol C (36)

Figure 3 Pentacyclic pyridoacridine alkaloids 27-81.

methylene singlet at dH 6.09/dC 77.2 characterized the benzoxathiole ring was observed. NOESY, ROESY, and NOE experiments may be used to assign the relative stereochemistry and conformation.

.. / \ O H3C CH3

Petrosamine (37) Debromopetrosamine trifluoroacetate salt

O NH * JL

H O CF3

O NH - • -CF

■■ " - CF3

Ecionine A (39)

Ecionine B (40)

Cystodimine A (41)

Cystodimine B (42)

Shermilamine A (43) Debromoshermilamine A = Shermilamine

B (44)

N-Deacetylshermilamine B (45) Shermilamine C (46)

Fig. 3 (continued)

Shermilamine D (47)

Shermilamine F (49)

NO(Me)2

Shermilamine E (48)

Sebastianine A (50)

Lissoclinidine trifluoroacetate salt (51) Lissoclinidine B (52)

Labuanine A (53)

Ascididemin (54) "N

3-Hydroxyascididemin (55) 8,9-Dihydro-11-hydroxyascididemin (56)

11-Hydroxyascididemin (57)

12-Deoxyascididemin (58) Fig. 3 (continued)

9-Hydroxyisoascididemin (59)

Meridine (60)

Meridin-12(13H)-one (61)

9-Aminobenzo[è]pyrido[4,3,2-rfe][1,10]phenanthrolin-8(8H)-one (62) HO„

Ancorine A (63)

Cnemidine A (64)

Arnoamine A (65)

Arnoamine B (66)

Arnoamine C (67)

Arnoamine D (68)

Fig. 3 (continued)

DMSO-<i

DMSO-d

Demetyldeoxyamphimedine (75)

1 -Hydroxy-deoxyamphimedine trifluoroacetate salt (76)

3-Hydroxy-deoxyamphimedine trifluoroacetate salt (77)

Neoamphimedine (78)

Deoxyamphimedine (79)

Cystodamine (80)

2-Bromoleptoclinidinone (81) Fig. 3 (continued)

2.4. Mass spectroscopy (MS)

Pyridoacridines structures were established by extensive spec-troscopic analysis. Low and high resolution mass spectro-scopic data (EI, FAB, and ESI-MS) were used to establish the molecular formula. Few fragments corresponding to the loss of CHO, CO, CH, and HCN were observed.31 The imino-quinone system as in cytodytin A (3) is easily reduced in the ionization stage of a mass spectrometer giving (MH + 2) in FABMS and (M + 2) ion for EIMS, identical for quinines.20 The compound containing one bromine atom will have two peaks in the molecular ion region in an approximately 1:1 ratio. Additionally, isotopic cluster of peaks (ratio 1:2:1) is typical for the presence of two bromines in the structure.32 In the mass spectrum, the appearance of a peak for [M-S2 + H]+ confirmed the sulfur ring system.33 Also, the mass peak at m/z 58 [N(CH3)2] indicated the presence of dimethy-lamino functionality.41

2.5. Stereochemistry determination

The X-ray analysis of suitable crystals is another technique for the assignment of the absolute configuration of the stereocen-

Nordehydrocyclodercitin (82)

N—CH3

Stellettamine (83)

Cycloshermilamine D (84) 13-Didemethylaminocycloshermilamine D

I OH H

Sebastianine B (86)

(Stl^RiX^ I СНэ|

Segoline A 87)

HN—\

Isosegoline A (88)

Segoline B (89)

H^Jz-fST CH3

Segoline C (90)

Cyclodercitin (91)

Figure 4 Hexacyclic pyridoacridine alkaloids 82-91.

ters present in the molecule besides being a valuable way to confirm the constitution and finally the conformation of the natural product. In many cases, this technique cannot be used since crystals of suitable quality are not usually available. Therefore, after finishing all of the abovementioned spectro-scopic investigations, a crystal-structure analysis can be carried out. Also, the configuration was deduced by comparison of experimental circular dichroism (CD) spectra with calculated spectra.

Eilatin (92)

Eudistone A (94)

Eudistone B (95)

Figure 5 Heptacyclic 92 and octacyclic 93-95 pyridoacridine alkaloids.

Conflict of interest

We have no conflict of interest to declare.

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